Whey Protein Composition with a Reduced Astringency

ABSTRACT

The invention relates to a sterilized liquid or semi-solid acid enteral composition comprising per 100 ml 9 to 20 g of non-hydrolysed globular proteins, fat and at least 100 mg of divalent metal cations and having a pH ranging between 3 and 5. 
     The invention further relates to a method for preparing a composition according to the invention, comprising a step wherein at least the non-hydrolysed globular proteins are subjected to a direct steam injection (DSI) at specific holding values, such as a holding temperature of 100 to 140° C. during a holding time of about 0.5 to 10 seconds, followed by a homogenization step and a sterilization step 
     The composition according to the invention has a reduced astringency and can be used for medical purposes, such as for stimulating muscle protein synthesis in a mammal, in particular for treating sarcopenia, and for specific groups of people, such as elderly and sportsman.

FIELD OF THE INVENTION

The present invention relates to a sterilized liquid or semi-solid acidenteral composition comprising a high amount of non-hydrolysed globularproteins, such as whey, fat, and a high amount of divalent metalcations, such as calcium and magnesium, and having a reducedastringency, methods for the preparation of such composition and use ofsuch composition in the manufacture of a nutritional food, to be used asa complete food or as a nutritional supplement. The present inventionfurther relates to a nutritional composition useful for medicalpurposes, such as for stimulating muscle protein synthesis in an mammal,in particular for treating sarcopenia, and for specific groups ofpeople, such as elderly and sportsman.

BACKGROUND OF THE INVENTION

Some subjects need nutrition, either as a supplement, or as a completenutrition, in the smallest volume of liquid, that is still effective forits intended purpose.

These subjects can be cachectic patients or persons suffering fromend-stage AIDS, cancer or cancer treatment, severe pulmonary diseaseslike COPD (chronic obstructive pulmonary disease), tuberculosis andother infection diseases or persons that experienced severe surgery ortrauma like burns. Furthermore, persons suffering from disorders in thethroat or mouth such as oesophageal cancer or stomatitis and personshaving problems with swallowing like dysphagic persons, require specialliquid, low-volume nutrition. Also, persons just suffering from reducedappetite or loss of taste, will benefit from low-volume, preferablyliquid, food.

These subjects can also be elderly persons, in particular frail elderlyand elderly at risk of becoming frail. In this regard, although anelderly person's energy needs may be reduced, their ability to consumeproducts may also be diminished. For example, they may have difficultyconsuming a product due to, e.g., swallowing difficulties, or due thetoo large amount of product they need to consume to meet the dailyintake of nutrients. Hence, compliance is not optimal, and often, theintake is suboptimal, leading to suboptimal nourishment, and in the end,to malnutrition.

These subjects can also be sportsmen (male or female), as a sportsmanalso may benefit from a concentrated protein drink.

Due to a prerequisite of at least six months of shelf life in general,preferably at least 12 months, whey protein compositions need to undergosome sort of sterilization treatment in order to reduce the number of orremove possible pathogens, for instance spores, bacteria and othermicro-organisms, which cause spoilage of the protein composition,preferably by using heat (sterilization, pasteurization), radiation(UV-treatment), or filtration methods (ultrafiltration, diafiltration,nanofiltration). Preferred sterilization treatments include heattreatments at high temperatures for a short period, such as using a UHT(Ultra High Temperature) treatment. However, when subjecting wheyproteins to heat, whey proteins are rapidly denaturated whereby the wheyprotein globular structure enfolds, and at a pH between 3 and 7 may formagglomerates and macrostructures, which are visible as a haze orturbidity. Eventually, the agglomerates will sediment and thenutritional composition will become unacceptable for furtherconsumption. The use of acid whey (i.e. whey with a pH<7, preferablywith a pH between 3 and 5), either obtained from an acid whey process(also known as “sour whey”), or by acidification of whey (acidifiedwhey), obtained from acidifying sour whey or sweet whey by e.g. theaddition of an acid such as phosphoric acid, is preferred because acidwhey is less prone to pathogens and hence, only needs a mildsterilization treatment by heat, such as a pasteurization or UHTtreatment. Furthermore, an acid whey protein composition has a morepreferred taste and smell than a neutral (pH about 7) whey proteincomposition.

The aforementioned groups of subjects may be sensitive to foodconsistency and to the organoleptic properties of an acid compositioncomprising a high amount of non-hydrolysed globular proteins, such aswhey proteins, such as, for instance viscosity, taste, smell, colour andmouth feel, in particular astringency.

Acid whey protein solutions elicit an astringent taste sensation in themouth, irrespective of the source of whey (WPI, WPC and others).Although the exact mechanism of astringency by whey proteins is notknown, it has been published (Astringency of Bovine Milk Whey Protein,H. Sano, T. Egashira, Y. Kinekawa, and N. Kitabatake, J. Dairy Sci.88:2312-2317) that most of the whey protein precipitates in the mouth atabout pH 5. When an acid WPI solution (pH 3.5) is placed in the oralcavity, the acid solution is mixed with saliva (pH of about 7), causingthe pH of the whey protein solution to increase but to remain at a pH<5.At this pH (near the iso-electric point of the whey protein), wheyprotein would precipitate in the mouth. This precipitate is formed inthe oral cavity and would induce astringency in a similar way to thecomplex precipitation formed by salivary protein and polyphenoliccompounds, as can be found in wine, green tea and some fruits.

Furthermore, it was established that astringency increases withincreasing whey concentrations and shows a maximum at pH 3. This makesastringency to become a real taste problem in nutritional compositionshaving a high amount of whey and an acidic pH. The problem is inparticular apparent at an acid pH of about 3, in particular at a pHbetween 3 and 5.

It is further contemplated that the presence of divalent metal cations,such as magnesium and calcium—both important nutrients—may contribute toastringency. Further, it is contemplated that the presence of divalentmetal cations, in particular calcium may adversely affect the solubilityof whey protein and/or adversely affect viscosity of a liquid comprisingwhey protein, and/or adversely affect the shelf life, in particular incase the whey protein concentration is relatively high, or the liquid isheat-treated.

In view of expected problems with respect to organoleptic properties, inparticular astringency, protein solubility and/or controlling viscosity,the skilled person would therefore not consider to provide a sterilizedliquid or semi-solid acid enteral composition comprising a high amountof non-hydrolysed globular proteins, such as whey, fat, and a highamount of divalent metal cation, as he would not expect to be able toprovide such as product with satisfactory properties for the consumer.

Therefore, a problem underlying the present invention is how to providea sterilized liquid or semi-solid acid enteral composition comprising ahigh amount of non-hydrolysed globular proteins, such as whey, fat, anda high amount of divalent metal cations, such as calcium and magnesium,and having satisfactory properties, in particular satisfactory shelflife and satisfactory organoleptic properties, for providing nutrition,either as a supplement, or as a complete nutrition.

In particular, a problem underlying the invention is how to provide sucha product with a satisfactory shelf life and no or a low astringencyand/or no or a low sandiness.

More in particular, a problem underlying the invention is how to providea sterilized liquid or semi-solid acid enteral composition comprising ahigh amount of non-hydrolysed globular proteins, such as whey, fat, anda high amount of divalent metal cation in a relatively small volume ofliquid, whilst supporting nutrition and well-being in the differentsubject groups mentioned above.

The inventors have now found that such a problem is solved by preparinga sterilized liquid or semi-solid acid enteral composition in a specificway, namely using a method for producing said composition, comprising atleast a direct steam injection (DSI) step, whereby the DSI is used fornon-sterilizing purposes. Thus, the present invention makes it possibleto provide the specific composition as defined herein below and in theclaims, as an industrially applicable composition.

PRIOR ART FOR THE INVENTION

Major technical difficulties exist in producing a sterilized liquid orsemi-solid acid enteral composition comprising a high amount ofnon-hydrolysed globular proteins, such as whey, fat, and a high amountof divalent metal cations, such as calcium and magnesium, and having areduced astringency.

EP 1 894 477 A1 (Nestec S. A., 5 Mar. 2008) discloses the formation of acoated denaturated supramolecular protein core structure (aliposome-like structure) comprising a whey protein aggregate and alipidic bilayer (sulfated butyl oleate) for reduction of the astringencyof protein supramolecular structures (in particular micelles).

JP 57189657 A (Mitsubishi, 22 Nov. 1982) discloses a soya milk drink,free of astringency, by adding a fatty acid ester to soya milk,homogenizing the mixture, and heat treating at ≧70° C.

WO 2009/112036 (Arla Foods, 17 Sep. 2009) discloses whey proteinbeverages with a reduced astringency comprising 0.5-15 weight % of wheyand a shielding agent, in particular a monoglyceride.

WO 2007/108827 (Novartis, 27 Sep. 2007) discloses the use of DSI forreducing the viscosity of a milk protein isolate composition.

Nutritional compositions with a high amount of non-hydrolysed globularprotein, in particular whey, have been described, e.g. in WO 2009/113858(NV Nutricia, 17 Sep. 2009), in WO 2009/072884 (Nutricia, 11 Jun. 2009).

WO 2010/043415 (Nestec SA, 22 Apr. 2010) discloses a shelf-stable acidwhey composition comprising 10.67 g/100 g of WPI and 5.64 g/100 g of WPH(hydrolysate)—Example 3. DSI is used as a sterilization treatment (120°C./11 sec, flash 80° C.). It is not mentioned to include divalent metalcations. The presence of a whey hydrolysate gives the resultingcomposition a bad taste.

SUMMARY OF THE INVENTION

The inventors have now established that a sterilized liquid orsemi-solid acid enteral composition comprising a high amount ofnon-hydrolysed globular proteins, such as whey, fat, and a high amountof divalent metal cations, such as calcium and magnesium, and havingsatisfactory organoleptic properties, in particular a reducedastringency, is obtained using a process which includes a step wherein acomposition is subjected to a direct steam injection (DSI) step atspecific holding values and in a specific combination with other processsteps, in particular after the step of homogenisation said compositionand before a final sterilization treatment (meaning either sterilizationor pasteurization). In a preferred embodiment, the invention provides asterilized liquid or semi-solid acid enteral nutritional compositioncomprising per 100 ml of the composition 9 to 20 g of non-hydrolysedglobular protein, fat, and at least 100 mg of divalent metal cations,having a pH ranging between 3 and 5, preferably ranging between about3.7 and about 4.3, more preferably a pH of about 3.8, about 3.9, about4.0, about 4.1 or about 4.2, most preferably a pH of about 4.0.” In anadvantageous embodiment, such a composition is highly appreciatedbecause of its low astringency or the absence of astringency and/orbecause of its low sandiness or absence of sandiness.

In a further embodiment, the invention provides a sterilized liquid orsemi-solid acid enteral nutritional composition according to theinvention further comprising one or more of carbohydrates and dietaryfibres. Such a composition is useful and applicable for medicalpurposes, such as for sarcopenia, and for specific groups of people,such as elderly and sportsman.

In a further embodiment, the invention provides the use of saidsterilized liquid or semi-solid acid enteral nutritional compositionaccording to the invention for the manufacture of a nutritionalcomposition for providing nutrition to a person in need thereof.

In a further embodiment, the invention provides a method for thepreparation of a sterilized liquid or semi-solid acid enteralcomposition comprising per 100 ml of said composition 9 to 20 g ofnon-hydrolysed globular proteins, fat, and at least 100 mg of divalentmetal cations and having a pH ranging between 3 and 5, comprising a stepwherein at least the non-hydrolysed globular proteins are subjected to adirect steam injection (DSI) at specific holding values, such as aholding temperature of 100 to 140° C. during a holding time of about 0.5to 10 seconds, preceded by a homogenization step. Such a method is avaluable process tool in obtaining whey-based compositions comprisingper 100 ml of said composition a high whey concentration, in particularbetween 9 and 20 g, fat and divalent metal cations.

In a further embodiment, the invention provides a method for thepreparation of a sterilized liquid or semi-solid acid enteralcomposition according to the invention, comprising the consecutive stepsof:

-   -   a) preparing an aqueous solution comprising amounts of divalent        metal cations, in particular calcium and magnesium,        non-hydrolysed globular proteins and fat, such that said        sterilized liquid or semi-solid acid enteral composition        comprises per 100 ml of said composition 9 to 20 g of        non-hydrolysed globular proteins, fat and at least 100 mg of        divalent metal cations, and having a pH ranging between 3 and 5;    -   b) homogenizing the resulting solution essentially obtained by        step a); and    -   c) subjecting the resulting solution essentially obtained by        step b) to a direct steam injection process at a holding        temperature of 100 to 140° C. during a holding time of about 0.5        to 10 seconds.

In a further embodiment the invention provides a liquid or semi-solidacid enteral nutritional composition obtained or obtainable by a methodaccording to the invention. Such a product is in particularcharacterized by a relatively low astringency and/or sandiness, comparedto a product having a comparative ingredient composition that has beenobtained using a conventional technique or in a method wherein DSI isperformed prior to homogenization.

DETAILED DESCRIPTION OF THE INVENTION Definitions

Within the context of the present invention, an elderly person is aperson of the age of 50 or more, in particular of the age of 55 or more,more in particular of the age of 60 or more, more in particular of theage of 65 or more. This rather broad definition takes into account thefact that the average age varies between different populations, ondifferent continents, etc. Most developed world countries have acceptedthe chronological age of 65 years as a definition of ‘elderly’ or olderperson (associated with the age at which one may begin to receivepension benefits), but like many westernized concepts, this does notadapt well to e.g. the situation in Africa. At the moment, there is noUnited Nations (UN) standard numerical criterion, but the UN agreedcut-off is 60+ years to refer to the older population in Western world.The more traditional African definitions of an elder or ‘elderly’ personcorrelate with the chronological ages of 50 to 65 years, depending onthe setting, the region and the country.

Within the context of the present invention, enteral means any form ofadministration that involves any part of the gastrointestinal tract,i.e. by mouth (orally), by gastric feeding tube, duodenal feeding tube,or gastrostomy, and rectally, in particular by mouth (orally). Hence,when referring to an enteral composition, this means that thecomposition is suitable for enteral administration.

Within the context of the present invention, the term “astringency” isused for a puckering or mouth drying sensation, which appears after awhile in the mouth cavity after the consumption of a food. Thissensation “astringency” is also characterized by terms such as rough,dry, mouth coating, or filmy mouth-feel, suggesting finely dividedinsoluble particles in the mouth after consumption of a food. Hence,astringency is not a taste, but a physical mouth-feeling and timedepended feeling in the mouth cavity. In the same sense the term“non-astringent” is used, i.e. when no puckering or mouth dryingsensation is observed in the mouth cavity when consuming a food product,such as by a trained tasting panel, in a test procedure such as thefollowing. The “astringency value” may, as described in the Examplesbelow, be determined or measured by a trained tasting panel followingconventional specific sensory methods or by the analytical method suchas the “Saliva-Beverage Interaction test” as described WO 2009/112036.In the context of the present invention, the term “reduced astringency”is used to denote an astringence which is reduced, possibly to completeabsence of a noticeable astringency, in comparison to an acidcomposition comprising non-hydrolysed globular proteins, such as whey,having a high amount of protein and a high amount of calcium, butproduced with a method according to the state of the art.

The term “sandiness” also refers to a sensory property of a liquid orsemi-solid composition and typically relates to the presence of grainswhich causes an extraneous feeling remaining on the tongue as a distinctaftertaste. This property can be considered as an opposite to‘smoothness’ or ‘smooth mouth feel’ and is an important factor in theacceptance of liquid as well as semi-solid compositions.

Within the context of the present invention, the term “sterilizationtreatment” and the term “sterilization” is meant to comprise any methodusing heat (sterilization, pasteurization), radiation (UV-treatment),and/or filtration (ultrafiltration, diafiltration, nanofiltration) toreduce the number of or remove possible pathogens. Preferably, thesterilization treatment includes a heat treatment at a high temperaturefor a short period, such as a UHT (Ultra High Temperature) treatment.Hence, within the context of the present invention, pasteurization iscomprised within sterilization.

Within the context of the present invention, a “sterilized composition”is a composition that is obtained or obtainable by subjecting acomposition to a sterilization treatment. In general, the quantity ofpotentially pathogenic micro-organisms of the sterilized compositionmeets food safety requirements, as applicable e.g. in the US or EU. Inparticular, a sterilized composition in accordance with the inventionmaintains to meet such requirement, for at least 6 months, preferably atleast 12 months, when stored in a sealed packaging at ambienttemperature (20° C.).

Within the context of the invention, the pH is the pH as measurable witha pH electrode, calibrated at pH 4 and pH 7, at a temperature of 20° C.

Within the context of the invention, the viscosity is the viscosity asmeasurable using an Anton Paar Physica MCR301 rheometer with a CP50-1/PCcone (diameter 50 mm, 1° difference between middle and outside) at 20°C. at 100 s⁻¹.

Within the context of the invention, in general the shelf life of aproduct is the period, starting from its manufacture, during which theproduct remains suitable for consumption. In particular, during itsshelf-life, the product maintains an acceptable microbiological quality,maintains fluidity, a pH in the range of 3 to 5, 9 to 20 g per 100 ml ofnon-hydrolysed globular protein, fat and at least 100 mg of divalentmetal cations in the product per 100 ml of said product. In a preferredembodiment, the product maintains a viscosity of about 200 mPa·s orless, more preferably of 100 mPa·s or less during its shelf life.

The term “about” is in particular used herein to indicate a range of±10%, more in particular of ±5% around a given value.

Globular Proteins

The invention is generally concerned with globular proteins. Globularproteins may be single peptide chains, two peptide chains or morepeptide chains which interact in the usual ways. A globular protein mayhave portions of the chains with helical structures, pleated structures,or completely random structures. Globular proteins are relativelyspherical in shape as the name implies. In the art, globular proteinsare described as proteins of which the protein chain, including thesecondary structure elements, is tightly folded into a more or lessspherical shape (cf. Dairy Science and Technology, 2^(nd) ed. ISBN0-8247-2763-0). The tertiary structure assumed by a globular proteinmolecule tends to be such that the non-polar side chains are directedinward to allow interaction with one another and the polar side chainsare typically oriented outward such that they are exposed to adjacentpolar water molecules. A globular protein herein is to be understood asa protein which is globular in its non-denatured state. They aredistributed in both plant and animal tissues. For instance, albumins canbe found in blood (serum albumin), milk (lactalbumin), egg white(ovalbumin), lentils (legumelin), kidney beans (phaseolin), and wheat(leucosin). Globulins can be found in blood (serum globulins), muscle(myosin), potato (tuberin), Brazil nuts (excelsin), hemp (edestin), whey(lactoglobulins, immunoglobulins, and lactoferrins), pea and lentils(legumin, vicilin), and soy. Also, many enzymes and other vegetableproteins are globular proteins. More specifically, the invention isconcerned with globular protein selected from the group consisting ofwhey protein, pea protein, soy protein, and any mixture thereof, more inparticular with whey proteins.

When referred herein to a “non-hydrolysed” globular protein, this meansthat the protein is fully intact or only to a minor extent containshydrolysed fragments. A minor extent is an extent in which the globularnature of the protein is essentially maintained. The hydrolysedfragments—if present at all—in particular make up less than 10 weight %,such as, for instance 1 to 5 weight % relative to the total weight ofthe globular protein.

Hence, with the term globular protein is meant the collection ofproteins which are globular in nature, but may contain minute amounts ofhydrolysed fragments and/or uncoiled fragments.

The amount of non-hydrolysed globular protein, preferably whey, rangesbetween 9 and 20 g per 100 ml. Preferably, the amount of non-hydrolysedglobular protein, preferably whey, ranges between 9 and 16 g per 100 ml.Advantageously, the lower limit in the amount of non-hydrolysed globularprotein, preferably whey, is any of 9, 10, 11, 12, 13, 14, 15, 16, 17,18 or 19 g per 100 ml. Advantageously, the upper limit in the amount ofnon-hydrolysed globular protein, preferably whey, is any of 20, 19, 18,17, 16, 15, 14, 13, 12, 11 or 10 g per 100 ml. More preferably, theamount of non-hydrolysed globular protein, preferably whey, rangesbetween 9 and 20, or 9 and 18, or 9 and 16, or 9 and 14, or 9 and 12, or10 and 20, or 10 and 18, or 10 and 16, or 10 and 15, or 10 and 14, or 10and 12 g per 100 ml. In a specific embodiment, the amount ofnon-hydrolysed globular protein, preferably whey, is equal to about 10 gper 100 ml of the composition. Preferably, the amount of non-hydrolysedglobular protein is at least 85 weight % of the total proteinaceousmatter in the composition according to the invention, the rest of theproteinaceous matter being selected from the group comprising anon-globular protein, a hydrolysed protein, an oligopeptide, a peptideand a free amino acid.

In a specific embodiment, the non-globular protein is selected from thegroup of casein, caseinate, micellar casein isolate, and any mixturethereof.

In a specific embodiment, the free amino acid is selected from the groupof branched chain amino acids and salts thereof; in a particularembodiment, the free amino acid is L-leucine.

Whey Proteins

One of the most superior classes of food protein is whey protein. It hasan excellent amino acid profile for a purpose of the invention, highamount of cystein, rapid digestion, and interesting bioactive proteins(lactoglobulins, immunoglobulins, and lactoferrins). Nutritionallyspeaking, whey protein is known as a naturally complete protein becauseit contains all of the essential amino acids required in the daily diet.It is also one of the richest sources of branched chain amino acids(BCAAs, in particular leucine) which play an important role in muscleprotein synthesis. Moreover, some of the individual components of wheyprotein have been shown to prevent viral and bacterial infection andmodulate immunity in animals. Whey protein is the preferred choice ofproteins to treat persons suffering from sarcopenia, but is alsosuitable for healthy persons, such as sportsmen and (active) elderly.

As a source of whey protein to be used in the present invention, anycommercially available whey protein source may be used or any. wheyobtained by any process for the preparation of whey known in the art, aswell as whey protein fractions prepared thereof, or the proteins thatconstitute the bulk of the whey proteins being β-lactoglobulin,α-lactalbumin and serum albumin, such as liquid whey, or whey in powderform, such as whey protein isolate (WPI) or whey protein concentrate(WPC). Whey protein concentrate is rich in whey proteins, but alsocontains other components such as fat and lactose. Furthermore, wheyoriginating from sweet whey may contain glycomacroprotein (GMP), acaseine-related non-globular protein, which is also soluble at a pH atwhich the whey proteins are soluble and hence, difficult to separatetherefrom. Typically, whey protein concentrate is produced by membranefiltration. On the other hand, whey protein isolate consists primarilyof whey proteins with minimal amounts of fat and lactose. Whey proteinisolate usually requires a more rigorous separation process such as acombination of microfiltration and ultra-filtration or ion exchangechromatography. It is generally understood that a whey protein isolaterefers to a mixture in which at least 90 weight % of the solids are wheyproteins. A whey protein concentrate is understood as having apercentage of whey proteins between the initial amount in the by-product(about 12 weight %) and a whey protein isolate. In particular, sweetwhey, obtained as a by-product in the manufacturing of cheese, acidwhey, obtained as a by-product in the manufacturing of acid casein,native whey, obtained by milk microfiltration or rennet whey, obtainedas a by-product in the manufacturing of rennet casein, may be used aloneor in combination as source of globular whey proteins.

Furthermore, whey proteins may originate from all kinds of mammaliananimal species, such as, for instance cows, sheep, goats, horses,buffalo's, and camels. Preferably, the whey protein is of bovine origin.

Preferably, the whey protein source, used for preparing a productaccording to the invention, is available as a powder, preferably thewhey protein source is selected from the group consisting of wheyprotein concentrate (WPC), whey protein isolate (WPI), and any mixturethereof.

According to a specific embodiment, the whey is a mixture ofnon-acidified (i.e. neutral) WPI and acidified WPI. Amounts of acidifiedand non-acidified WPI may range between 10 weight % and 90 weight %,such that any weight ratio is obtained between 10/90 and 90/10.Preferably, the weight ratio acidified:neutral whey is in the range of50:50 to 70:30. A weight ratio acidified:neutral whey of about 60:40 isparticularly preferred.

Whey protein isolate consists mainly of a mixture of β-lactoglobulin,α-lactalbumin and serum albumin, and optionally GMP if the whey sourceis sweet whey. The three first proteins are globular proteins that aresensitive to aggregation in the denaturated state. The denaturationtemperature of β-lactoglobulin is pH-dependent; at pH 6.7, irreversibledenaturation occurs when the protein is heated at temperatures aboveabout 65° C. In the denaturated state, a free thiol group is exposed.This free thiol group can initiate inter-protein disulfide interactionsleading to a polymerization reaction resulting in aggregate formation.Also two disulfide bridges, present in native β-lactoglobuline, areinvolved in the polymerization reaction and also other sulphurcontaining groups including cysteine residues are thought to play arole.

α-Lactalbumin also has a denaturation temperature of about 65° C. Sinceα-lactalbumin does not have a free thiol group (only four disulfidebridges), solutions of pure α-lactalbumin are not irreversiblydenaturated under most processing conditions. However, in the presenceof β-lactoglobulin, as is the case in e.g. a whey protein concentrate orisolate, α-lactalbumin is more sensitive to irreversible denaturationthrough the formation of α-lactalbumin/β-lactoglobulin complexes inwhich also disulfide bridges in β-lactoglobuline and α-lactalbumin areinvolved via interchange reactions. Also, the circumstance thatα-lactalbumin contains cystein residues is considered to contribute to acertain sensitivity to irreversible denaturation.

Denaturated β-lactoglobulin and α-lactalbumin are both sensitive tocalcium; this is particularly the case in the pH range of about 5 toabout 8 where the protein carries a neutral to net negative charge. AtpH 4, the protein carries a net positive charge and is less (but still)sensitive to calcium-induced aggregation.

The size, shape and density of the protein aggregates in a matrix areinfluenced by a number of environmental and processing parametersincluding temperature, heating rate, pressure, shear, pH and ionicstrength and other ingredients in the matrix, such as, for instancecarbohydrates, minerals, acids, fat, etc. Depending on the combinationof these parameters and ingredients, the aggregates may form aspace-filling network (gel), fibrils or compact micro-particles. Forexample, microparticulated whey can be formed under specific ionicstrength and shear conditions. These particles have a compact structure,a high intrinsic viscosity and a low specific volume. Further, it isknown that a relationship exists between aggregates size and heatingtemperature for microparticulated whey produced under shear conditions.Microparticulated whey protein has received a lot of interest lately forapplication as a fat replacer or viscosity enhancer for yoghurt.

One of the major problems encountered with the production of liquidready-to-use compositions containing globular proteins in general, andwhey proteins in particular, is their limited processability andheat-sensitivity. As these proteins are heated above their denaturationtemperature in a sterilization process, they unfold and are transformedinto a reactive state, polymerize into aggregates or gels. As aconsequence, the heat-treated liquid composition exhibits unwantedsensorial attributes like chalkiness, sandiness, lumpiness. Besides, theshelf life of these products is limited in that sediment and/or creamlayers are formed soon after production or in that age-thickeningoccurs. In a composition with a high amount of globular protein, inparticular whey, these instabilities are even more pronounced and resultin products with an unwanted high viscosity and extensive fouling andblocking of the heating equipment.

Surprisingly, the inventors have now found that it is possible toprepare a sterilized liquid or semi-solid acid enteral nutritionalcomposition by means of a method wherein a composition that comprisesmainly globular proteins as a protein source, in particular wheyproteins, is subjected to a specific heat-treatment that comprises astep of subjecting the whey proteins to a DSI treatment at conditionswhich may be insufficient per se to sterilize or pasteurize thecomposition comprising the globular proteins, in particular wheyproteins.

Without being bound (or restricted) by theory, it is believed thatraising the temperature has a different effect on both denaturation andaggregation. While under a temperature of about 100° C., the rate ofaggregation is higher than the rate of denaturation, this behaviour isquickly reversed at temperature above about 100° C. At a temperaturebelow about 100° C., heating leads to the formation of long proteinstrains that may form disulfide bonds and aggregate and form largeparticles that eventually sedimentate. At a temperature above about 100°C., globular proteins quickly start to denaturate. Hence, a slowheat-treatment just above the denaturation temperature of the whey leadsto extensive polymerization and voluminous protein aggregates. Also,when the whey is heated to high temperatures (i.e. far above the proteindenaturation temperature, for example at about 110° C.) via a slowheating process, i.e. a process in which the temperature of the proteinsolution is raised gradually, for example 0.1 to 2° C. per second, usinge.g. retort, plate or tubular heat exchangers, the whey exhibitsextensive polymerization during heating up when process temperaturespass the temperature window just above the denaturation temperature ofthe whey protein. As a result, the product is too thick, lumpy, sandyand extensive fouling is observed in the heating apparatus, inparticular when high amounts of calcium are present per 100 ml ofcomposition, such as, for instance more than 100 mg, more in particularmore than 200 mg in compositions comprising 9 to 20 gram of globularprotein, in particular whey protein.

Using the method according to the invention, by quickly and shortlyheating the globular proteins well above the denaturation temperature ofthe whey protein, the thiol group of β-lactoglobulin, the mainconstituent of whey protein, is very quickly being exposed andtermination reactions forming disulfide bridges dominate initially afterheating. As a result, small, compact whey protein particles are formedwhich are largely inert in any further heat-treatment. Hence,surprisingly, it was found that the time for whey proteins to be spentin a temperature window just above the denaturation temperature, shouldbe minimized.

Surprisingly, as a result of said treatment, a resulting sterilizedliquid or semi-solid acid enteral nutritional composition has a longshelf life, typically at least 6 months, preferably 12 months or more,satisfactory organoleptic properties, such as no or a low astringencyand/or no or a low sandiness compared to a prior art sterilized liquidor semi-solid acid enteral nutritional composition.

Stabilizing Polysaccharide

In a preferred embodiment of this invention, the sterilized liquid orsemi-solid acid enteral nutritional composition also comprises apolysaccharide capable of stabilizing the small compact proteinparticles that are formed in the heat-treatment. These polysaccharidesare also referred to herein as “stabilizing polysaccharides”.

Without limiting the scope of the invention, it is hypothesized thatcertain polysaccharides having both positively and negatively chargedgroups or regions at a pH below the isoelectric point (IEP) of wheyprotein. At such a pH the protein will have a net positive charge, whichis assumed to interact with the negatively charged groups of thepolysaccharide. This interaction results in the protein particle beingsurrounded by the large polysaccharide molecules decreasing thelikelihood of protein particles coming within close distance of eachother and aggregate. Furthermore, it is hypothesized that the longpolysaccharide chains can form networks within the liquid matrixpreventing sedimentation of the protein particles.

Hence, in a preferred embodiment the stabilizing polysaccharide is apolysaccharide having positively as well as negatively charged groups ata pH within the range of 3-5, e.g. at a pH of 4.2. Furthermore, it ispreferred that the stabilizing polysaccharide does not interact with thecalcium ions to form firm gel structures. Polysaccharides that maysuitably be used for the purposes of the present invention include highmethoxy pectin and carboxymethyl cellulose. Preferred examples ofcarboxymethyl celluloses that may suitably be used in accordance withthe invention, include Clear+Stable 30 PA, Clear+Stable 100 PA andClear+Stable 2000 PA (Dow chemical). The term “high methoxy pectin”herein is to be understood as a methoxy pectin wherein at least 50% ofthe galacturonic acid groups are esterified with a methyl group.

In one embodiment of the invention, the stabilizing polysaccharide ishigh methoxy pectin, which is typically used in a concentration of0.01-1% (w/v), preferably 0.02-0.5% (w/v), more preferably 0.05-2%(w/v), for example in a concentration of 0.1% (w/v).

In another embodiment of the invention, the stabilizing polysaccharideis carboxy methyl cellulose, which is typically used in a concentrationof 0.1-10% (w/v), preferably 0.2-5% (w/v), more preferably 0.5-3% (w/v),most preferably in a concentration of 1-2% (w/v).

Direct Steam Injection

Direct Steam Injection (DSI) involves the discharge of steam (water at atemperature above 100° C.) into a liquid with a lower temperature thanthe steam. The steam condenses and gives up its heat to the surroundingliquid. As heat is transferred by direct contact between the steam andthe liquid, consequently this method is only used when dilution and anincrease in liquid mass is acceptable. Therefore, the liquid beingheated is usually water or an aqueous composition, such as a nutritionalcomposition. Furthermore, after flash-cooling of the heated liquid, mostof the added steam is lost again by evaporation under vacuum. DSI isused in the food industry since the early 1930s for use as asterilization treatment, and its principles are known to the skilledperson and will not be further disclosed herein in detail. Commercialapparatus can be bought e.g. from the company Tetra Pak ProcessingSystems BV, Houten, The Netherlands.

In this application, DSI is applied for non-sterilizing purposes, mostlyat non-sterilizing conditions. An overview of the DSI conditionsaccording to this application, in comparison with the conditions for UHTtreatment is shown in FIG. 1.

Recently, the use of DSI for non-sterilizing purposes has been disclosedin WO2007/108827 (Abbott) for the reduction of the viscosity of a highenergy (225-325 kcal/ml) milk protein isolate composition (comprisingabout 20% of whey) for MPI amounts of between 6.7 and 12.6 g/100 ml(which corresponds to about 1.3 to 2.5 g whey per 100 ml of liquidcomposition. However, no examples were given, illustrating the claimedeffect, nor is disclosed that the effect is related to a reduction ofastringency or that is can be applied to predominantly whey-basedcompositions, such as comprising 9 to 20 g/100 ml of whey proteins.

Method of Preparation of the Composition

The invention provides a method for the preparation of a sterilizedliquid or semi-solid acid enteral composition comprising 9 to 20 g ofnon-hydrolysed globular proteins, fat and at least 100 mg of divalentmetal cations and having a pH ranging between 3 and 5, comprising a stepwherein at least the non-hydrolysed globular proteins are subjected to adirect steam injection (DSI) at specific holding values, such as aholding temperature of 100 to 140° C. during a holding time of about 0.5to 10 seconds, preceded by a homogenization step.

In a preferred embodiment of the invention, a process as defined aboveis provided wherein the step wherein at least the non-hydrolysedglobular protein is subjected to DSI treatment, is followed by asterilization step.

As will be understood by those skilled in the art, the DSI treatment asdescribed herein may result in the liquid or semi-solid becomingsterilized or ‘commercially sterile’ without performing a separatesterilization, depending on e.g. the conditions applied during the DSItreatment. Hence, embodiments wherein the process described above is notfollowed by a separate sterilization step are also within the scope ofthis invention.

In another preferred embodiment of the invention, a process as definedabove is provided wherein at least the non-hydrolysed globular proteinand a stabilizing polysaccharide are subjected to homogenizationfollowed by DSI treatment and, optionally, a sterilization step.

The invention also provides a method for the preparation of a sterilizedliquid or semi-solid acid enteral nutritional composition comprising per100 ml of said composition 9 to 20 g of non-hydrolysed globular protein,fat, and at least 100 mg of divalent metal cations and having a pHranging between 3 and 5, comprising the consecutive steps of:

-   a) preparing an aqueous solution comprising amounts of divalent    metal cations, in particular calcium and magnesium, non-hydrolysed    globular proteins, fat, and, optionally, the stabilizing    polysaccharides, such that said sterilized liquid or semi-solid acid    enteral composition comprises per 100 ml of said composition 9 to 20    g of non-hydrolysed globular proteins, fat, at least 100 mg of    divalent metal cations and, optionally the stabilizing    polysaccharide, and having a pH ranging between 3 and 5;-   b) homogenizing the resulting solution essentially obtained by step    a);-   c) subjecting the resulting solution essentially obtained by step b)    to a direct steam injection process at a holding temperature of 100    to 140° C. during a holding time of about 0.5 to 10 seconds; and,    optionally,-   d) subjecting the resulting solution essentially obtained by step c)    to a sterilization treatment.

With “consecutive” is meant that the order in which the steps areimplemented is: step a) followed by step b), followed by step c),followed by (optional) step d). Steps implementing other actions may beintermittently added to the sequence of steps a), b), c) and d), withthe proviso the order of the steps a), b), c) and (optionally) d) is notchanged. Typical steps that may be added are:

-   -   preparing other solutions;    -   dissolving other macro constituents of a nutritional composition        (e.g. carbohydrates, fibres);    -   dissolving other constituents such as minerals, amino acids,        etc.;    -   mixing;    -   preheating;    -   adjusting the pH;    -   flash-cooling.

In a particularly preferred embodiment of this invention, a method forthe preparation of a sterilized semi-solid acid enteral composition,also referred to as ‘spoonable composition’, is provided as described inany of the foregoing, comprising the additional step f) of adding athickener or gelling agent to the liquid prior to or after DSItreatment.

With “the resulting solution essentially obtained” is meant the solutionessentially resulting from a previous process step, with the provisothat the solution may contain other components as a consequence of anintermitting process step e), such as, but not limited to, the additionof other nutritional components.

A preferred process according to the present invention for thepreparation of a sterilized liquid or semi-solid acid enteralcomposition according to the invention, comprises the consecutive stepsof:

-   e1) dissolving an amount of non-hydrolysed globular proteins in a    first aqueous solution, such that said sterilized liquid or    semi-solid acid enteral composition obtained comprises per 100 ml of    said composition 9 to 20 g of non-hydrolysed globular proteins;-   e2) dissolving an amount of minerals comprising divalent metal    cations, in particular calcium and magnesium, in a second aqueous    solution, such that said sterilized liquid or semi-solid acid    enteral composition obtained comprises per 100 ml of said    composition at least 100 mg of divalent metal cations; preferably,    this is performed at a pH of about 4.3.-   e3) mixing the second aqueous solution comprising divalent metal    cations, in particular calcium and magnesium, with the first aqueous    solution comprising an amount of non-hydrolysed globular proteins    such that said sterilized liquid or semi-solid acid enteral    composition obtained comprises per 100 ml of said composition 9 to    20 g of non-hydrolysed globular proteins and at least 100 mg of    divalent metal cations;-   e4) adding an amount of fat, preferably a liquid fat, to the    resulting solution essentially obtained from step e3)-   b) homogenizing the resulting solution essentially obtained by step    e4);-   e5) preheating the resulting solution essentially obtained by step    b);-   c) subjecting the resulting solution essentially obtained by step    e5) to a direct steam injection process at a holding temperature of    100 to 140° C. during a holding time of about 0.5 to 10 seconds;-   e6) flash-cooling the resulting solution essentially obtained by    step c);-   e7) optionally, adjusting the pH of the resulting solution obtained    by step e6); and, optionally,-   d) subjecting the resulting solution essentially obtained by step    e7) to a sterilization treatment.

A preferred process according to the present invention for thepreparation of a sterilized liquid or semi-solid acid enteralcomposition comprising per 100 ml of said composition 9 to 20 g ofnon-hydrolysed globular proteins, fat, and carbohydrates according tothe invention, comprises the consecutive steps of:

-   e1) dissolving an amount of non-hydrolysed globular proteins and an    amount of carbohydrates in a first aqueous solution, such that said    sterilized liquid or semi-solid acid enteral composition obtained    comprises per 100 ml of said composition 9 to 20 g of non-hydrolysed    globular proteins;-   e2) dissolving an amount of minerals comprising divalent metal    cations, in particular calcium and magnesium, in a second aqueous    solution, such that said sterilized liquid or semi-solid acid    enteral composition obtained comprises per 100 ml of said    composition at least 100 mg of divalent metal cations; preferably,    this is performed at a pH of about 4.3.-   e3) mixing the second aqueous solution comprising divalent metal    cations, in particular calcium and magnesium, with the first aqueous    solution comprising an amount of non-hydrolysed globular proteins    and an amount of carbohydrates such that said sterilized liquid or    semi-solid acid enteral composition obtained comprises per 100 ml of    said composition 9 to 20 g of non-hydrolysed globular proteins and    at least 100 mg of divalent metal cations;-   e4) adding an amount of fat, preferably a liquid fat, to the    resulting solution essentially obtained by step a);-   b) homogenizing the resulting solution essentially obtained by step    e4);-   e5) preheating the resulting solution essentially obtained by step    b);-   c) subjecting the resulting solution essentially obtained by step    e5) to a direct steam injection process at a holding temperature of    100 to 140° C. during a holding time of about 0.5 to 10 seconds;-   e6) flash-cooling the resulting solution essentially obtained by    step c);-   e7) optionally, adjusting the pH of the resulting solution obtained    by step e6); and, optionally,-   d) subjecting the resulting solution essentially obtained by step    e7) to a sterilization treatment.

Another preferred process according to the present invention for thepreparation of a sterilized liquid or semi-solid acid enteralcomposition according to the invention, comprises the consecutive stepsof:

-   e1) dissolving an amount of non-hydrolysed globular proteins and a    stabilizing polysaccharide in a first aqueous solution, such that    said sterilized liquid or semi-solid acid enteral composition    obtained comprises per 100 ml of said composition 9 to 20 g of    non-hydrolysed globular proteins;-   e2) dissolving an amount of minerals comprising divalent metal    cations, in particular calcium and magnesium, in a second aqueous    solution, such that said sterilized liquid or semi-solid acid    enteral composition obtained comprises per 100 ml of said    composition at least 100 mg of divalent metal cations; preferably,    this is performed at a pH of about 4.3.-   e3) mixing the second aqueous solution comprising divalent metal    cations, in particular calcium and magnesium, with the first aqueous    solution comprising an amount of non-hydrolysed globular proteins    such that said sterilized liquid or semi-solid acid enteral    composition obtained comprises per 100 ml of said composition 9 to    20 g of non-hydrolysed globular proteins and at least 100 mg of    divalent metal cations;-   e4) adding an amount of fat, preferably a liquid fat, to the    resulting solution essentially obtained from step e3);-   b) homogenizing the resulting solution essentially obtained by step    e4);-   e5) preheating the resulting solution essentially obtained by step    b);-   c) subjecting the resulting solution essentially obtained by step    e5) to a direct steam injection process at a holding temperature of    100 to 140° C. during a holding time of about 0.5 to 10 seconds;-   e6) flash-cooling the resulting solution essentially obtained by    step c); and, optionally,-   e7) adjusting the pH of the resulting solution obtained by step e6).

Step e1) is preferably performed at 1 to 70° C., preferably at 20 to 55°C. At higher temperatures, less foaming is observed. The proteins aredissolved in a volume of an aqueous solution, preferably water such as,for instance demineralised water, demi-water or tap water, such that,after dilution with other solutions in subsequent steps, an end volumeis obtained such that this end volume comprises 9 to 20 g ofnon-hydrolysed globular proteins per 100 ml of sterilized liquid orsemi-solid acid enteral composition.

Step e2) is preferably performed at 1 to 90° C., preferably at 20 to 30°C. Preferably, this step is performed at a pH of about 4.3. At this pH,minerals (in the form of salts, hydroxides, etc.) dissolve most easily.The minerals are dissolved in a volume of an aqueous solution,preferably water such as, for instance demineralised water, demi-wateror tap water, such that, after dilution with other solutions insubsequent steps, an end volume is obtained such that this end volumecomprises at least 100 mg of non-hydrolysed globular proteins per 100 mlof sterilized liquid or semi-solid acid enteral composition.

Preferred embodiments of the invention provide any of the above definedprocesses, wherein the step e7) of adjusting the pH of the solutionobtained by step e6) is performed. Furthermore, in an embodiment it ispreferred that steps e7) and d) are both performed.

Step a) is preferably performed at 1 to 90° C., preferably at 20 to 30°C.

Step b) is preferably performed at 1 to 90° C., preferably at 60 to 70°C. Preferably, the mixture is pumped by a high pressure pump through anarrow opening, a valve. Due to the very narrow opening a high speed isintroduced. When the pressure is 60 MPa (600 Bar.) the maximal velocitywill be about 600 m/s. The potential energy will be transformed inkinetic energy resulting in an increase in temperature and heavyturbulence. The increase in temperature corresponds with P/4, sohomogenisation at 60 MPa results in a temperature rise of 15° C. Theheavy turbulence results in a disruption of the fat globules. Since theresidence time of the product in the valve is so short, an enormousenergy density is created (10¹¹-10¹² Wm⁻³). As a consequence of the highenergy density, the oil droplets are disrupted into smaller droplets.Since the energy dissipation is not constant, the droplets formed willvary in size, so a particle size distribution will be created. Thebetter the valve of the homogeniser, the smaller the particle sizedistribution. If a valve is not in perfect state, a wider particle sizedistribution will be produced giving bigger droplets which can createproduct problems such as creaming.

Other steps may be performed at such temperatures that can easily beselected by the skilled person, without any inventive activity,depending on e.g. the apparatus used.

Divalent Metal Cations

With the term “divalent metal cation” is meant any positive chargedmetal ion with a charge equal to two. In particular is meant the ions ofmagnesium (Mg²⁺), calcium (Ca²⁺), zinc (Zn²⁺) and iron (Fe²⁺),preferably calcium (Ca²⁺) as these ions appear at relative highconcentrations in nutritional compositions, in particular to comply withFSMP regulations. Preferably, the composition according the invention isa nutritionally complete composition.

In an embodiment of the present invention, the amount of divalent metalions is at least 100 mg of divalent metal cations per 100 ml ofcomposition.

Preferably, the amount of divalent metal ions ranges between 100 mg/100ml and 600 mg/100 ml and preferably between 200 mg/100 ml and 500 mg/100ml. In a specific embodiment, the amount of divalent cations is about270 mg/100 ml.

Preferably, the divalent metal cation is selected from the groupconsisting of Ca, Mg and any mixture thereof, preferably Ca.

Preferably, the amount of calcium ranges between 100 mg/100 ml and 600mg/100 ml and more preferably between 200 mg/100 ml and 500 mg/100 ml.In a further embodiment, the amount of calcium is about 250 mg/100 ml.

Preferably, the amount of magnesium ranges between 10 mg/100 ml and 100mg/100 ml and more preferably between 15 mg/100 ml and 70 mg/100 ml. Ina further embodiment, the amount of magnesium is about 19 mg/100 ml.

Fat

According to the invention, the present sterilized liquid or semi-solidacid enteral composition should comprise an amount of fat (i.e. lipid).The amount of fat may range between 5 and 95%, preferably between 10 and70%, more preferably between 15 and 65%, relative to the total energyamount of the composition.

With regard to the type of fat, a wide choice is possible, as long asthe fat is of food quality. In an advantageous embodiment, thecomposition comprises a fat which is liquid under ambient conditions,i.e. at room temperature and a pressure of 1 atm.

The fat may either be an animal fat or a vegetable fat or both. Althoughanimal fats such as lard or butter have essentially equal caloric andnutritional values and can be used interchangeably, vegetable oils arehighly preferred in the practice of the present invention due to theirreadily availability, liquid form, ease of formulation, absence ofcholesterol and lower concentration of saturated fatty acids. In afurther embodiment, the present composition comprises rapeseed oil, cornoil and/or sunflower oil.

The fat may include a source of medium chain fatty acids, such as mediumchain triglycerides (MCT, mainly 8 to 10 carbon atoms long), a source oflong chain fatty acids, such as long chain triglycerides (LCT, mainly atleast 18 carbon atoms long) and phospholipid-bound fatty acids such asphospholipid-bound EPA or DHA, or any combination of the two types ofsources. MCTs are beneficial because they are easily absorbed andmetabolized in a metabolicallystressed patient. Moreover, the use ofMCTs will reduce the risk of nutrient malabsorption. LCT sources, suchas canola oil, rapeseed oil, sunflower oil, soybean oil, olive oil,coconut oil, palm oil, linseed oil, marine oil or corn oil arebeneficial because it is known that LCTs may modulate the immuneresponse in the human body.

In a specific embodiment, the fat comprises 30 to 60 weight % of animal,algal or fungal fat, 40 to 70 weight % of vegetable fat and optionally 0to 20 weight % of MCTs based on total fat of the composition. The animalfat preferably comprises a low amount of milk fat, i.e. lower than 6weight %, especially lower than 3 weight % based on total fat. Inparticular, a mixture of corn oil, egg oil, and/or canola oil andspecific amounts of marine oil is used. Egg oils, fish oils and algaloils are a preferred source of non-vegetable fats. Especially forcompositions that are to be consumed orally, in order to preventformation of off-flavours and to decrease a fishy after-taste, it isrecommended to select ingredients that are relatively low indocosahexaenoic acid (DHA), i.e. less than 6 weight %, preferably lessthan 4 weight % based on total fat. Marine oils containing DHA arepreferably present in the composition according to the invention in anamount lower than 25 weight %, preferably lower than 15 weight % basedon total fat. On the other hand, inclusion of eicosapentaenoic acid(EPA) is highly desirable for obtaining the maximum health effect.Therefore, in another embodiment, the amount of EPA may range between 4weight % and 15 weight %, more preferably between 8 weight % and 13weight % based on total fat. The weight ratio EPA:DHA is advantageouslyat least 6:4, for example between 2:1 and 10:1. In yet anotherembodiment, the amount of EPA is very low, such as 0.1 to 1 weight %,preferably 0.3 weight % or 0.6 weight %, based on total fat.

Also, the sterilized liquid or semi-solid acid enteral compositionaccording to the invention may beneficially comprise an emulsifier. Inprinciple, any food-grade emulsifier may be present. Suitableemulsifiers are commonly known. Generally the emulsifier contributes tothe energy amount of the fat in said composition.

Digestible Carbohydrate

In a specific embodiment of the present invention, the sterilized liquidor semi-solid acid enteral composition according to the inventionfurther comprises a digestible carbohydrate. Preferably, the digestiblecarbohydrate provides between 20 to 60% of the total energy amount ofthe composition according to the invention. The digestible carbohydratemay comprise either simple or complex carbohydrates, or any mixturethereof. Suitable for use in the present invention are glucose,fructose, sucrose, lactose, trehalose, palatinose, corn syrup, malt,maltose, isomaltose, partially hydrolysed corn starch, maltodextrins,glucose oligo- and poly-saccharides.

The composition of the digestible carbohydrate preferably is such thathigh viscosities, excessive sweetness, excessive browning (Maillardreactions) and excessive osmolarities are avoided. Acceptableviscosities and osmolarities may be achieved by adjusting the averagechain length (average degree of polymerisation, DP) of the digestiblecarbohydrates between 1.5 and 6, preferably between 1.8 and 4. In orderto avoid excessive sweetness, the total level of sucrose and fructose ispreferably less than 60%, more preferably less than 52%, more preferablyless than 40% of the weight of the carbohydrate, especially of thedigestible carbohydrate. Long-chain digestible carbohydrates such asstarch, starch fractions and mild starch hydrolysates (DE≧6, DE<20), mayalso be present, preferably in an amount of less than 25 weight %,especially less than 15 weight % of the digestible carbohydrate, andless than 6 g/100 ml, preferably less than 4 g/100 ml of the totalliquid enteral composition according to the invention.

Vitamins, Minerals and Trace Elements

The sterilized liquid or semi-solid acid enteral composition accordingto the invention may also contain a variety of vitamins, minerals andtrace elements.

In one embodiment of the present invention, the sterilized liquid orsemi-solid acid enteral composition according to the invention providesall necessary vitamins, most of the minerals and trace elements. Forexample, the composition according to the invention preferably providesabout 1.1 mg of zinc per 100 ml of the composition which is beneficialfor tissue repair in a healing patient. Preferably, the compositionaccording to the invention provides 16 mg of vitamin C per 100 ml of thecomposition to aid patients with more severe healing requirements.Further, preferably, the composition according to the invention provides1.2 mg iron per 100 ml of the composition. Iron is beneficial inmaintaining bodily fluids as well as circulatory system functions in anelderly patient.

The phosphorus amount may be above 10 mg per g of protein, and mayamount to, for instance, 125 mg/100 ml of total composition with acalcium to phosphorus weight ratio between about 1 and about 3. In afurther embodiment, the ratio is about 2.

Other ingredients may be present, such as vitamin A, carotenoids,vitamin D3, vitamin E, vitamin K, thiamin, riboflavin, niacin,panthotenic acid, vitamin B6, folic acid, vitamin B12, biotin, vitaminC, choline, lecithine and trace elements such as copper, manganese,selenium, molybdenum, chromium en iodine.

Thickener/Gelling Agent

As stated before, in an embodiment of the present invention, a liquidcomposition according to the invention may be used as the basis for themanufacturing of a semi-solid nutritional composition, also referred toas ‘spoonable composition’, such as a crème, a pudding, a custard, asoup, an ice cream, or a jelly. To this end, a liquid compositionaccording to the invention is processed to convert the low viscositycomposition according to the invention into a more solid or viscous one,e.g. by adding thickeners or gelling agents and further process themixture into the final semi-solid product, e.g. by subjecting it to aheat-treatment. Thickeners and/or gelling agents can also be present inthe formulation from an earlier stage of the process, or even dissolvedtogether with the nutrients at the beginning of the process. Hence,according to one embodiment, the invention is related to a semi-solidenteral nutritional composition obtainable from a nutritionalcomposition comprising per 100 ml of said composition 9 to 20 g ofnon-hydrolysed globular protein, fat, and at least 100 mg of divalentmetal cations and having a pH ranging between 3 and 5, by combining witha thickener or gelling agent.

Many conventional thickeners and/or gelling agents can be used inaccordance with the invention, including various gums, such as locustbean gum, guar gum, xanthan gum, gum arabic, carob gum, etc.; alginates;agar; carrageenan; cellulose and cellulose derivatives; starches,including modified starches, pectins, etc. In a preferred embodiment ofthe invention starch or pectin is used as thickener or gelling agent.

In one embodiment of the invention, the thickener or gelling agent ispectin, which is typically used in a concentration of 0.05-5% (w/v),preferably 0.1-1% (w/v), more preferably 0.2-0.5% (w/v), for example ina concentration of 0.35% (w/v).

In another embodiment of the invention, the thickener or gelling agentis starch, which is typically used in a concentration of 0.1-10% (w/v),preferably 0.2-5% (w/v), more preferably 0.5-2% (w/v), for example in aconcentration of 1% (w/v). In another embodiment the thickener isapplied in amounts sufficient to give the product the desired viscosityand/or structure. Preferably the viscosity of a semi-solid product, alsoreferred to herein as a spoonable product, is within the range between200-2500 mPa·s, preferably 300-2000 mPa·s, more preferably 400-1000mPa·s, all measured at 20° C. at a shear rate of 100 s⁻¹. Such products,meaning also spoonable products, are products that can readily beconsumed by spooning up the product from a container or plate.

In accordance with the present invention, liquid products are typicallyproducts that are pourable (at 20° C.), in particular pourable from anopened container in which they are contained. In particular, a productis liquid or pourable if its viscosity, as measured using the method asdescribed in the definitions section, lies below 200 mPa·s.

In another preferred embodiment of the invention the spoonable producthas the form of a hydrogel.

In one embodiment of the invention the composition comprises apolysaccharide that functions as a thickener as well as a stabilizer.For example, high methoxy pectin may be used in amounts sufficient toconvert the product into a more viscous or more solid product, which atthe same time will aid in the stabilization of the protein particlestypically through the mechanisms explained herein before. In aparticularly preferred embodiment the semi-solid composition comprises acombination of high methoxy pectin and a starch, typically in amountssufficient to impart the desired viscosity and to confer the stabilizingaction described herein before.

Nutritional Compositions

According to a preferred embodiment, the liquid enteral nutritionalcomposition according to comprises:

-   a) about 10 g of non-hydrolysed whey per 100 ml of the composition,    said protein providing about 56% of the total energy amount of the    composition;-   b) fat providing about 18% of the total energy amount of the    composition;-   c) optionally carbohydrate providing about 23% of the total energy    amount of the composition,-   d) about 250 mg per 100 ml of Ca and about 19 mg per 100 ml of Mg;    and-   e) having a pH of about 4.

Medical Use

The nutritional composition according to the invention canadvantageously be used for the nutritional management of a person inneed thereof, in particular wherein the person is an elderly person, aperson that is in a disease state, a person that is recovering from adisease state, a person that is malnourished, a sportsman, or an activeelderly. The nutritional composition according to the invention canadvantageously be used for the prevention or treatment of a disease orcondition involving muscle decline in a mammal. Alternatively, thenutritional composition according to invention can advantageously beused for the prevention or treatment of a disease or condition selectedfrom the group of sarcopenia, muscle loss, insufficient muscle proteinsynthesis, muscle degradation, muscle proteolysis, muscle atrophy,muscle dystrophy, muscle catabolism, muscle wasting, loss of musclestrength, loss of muscle mass, loss of muscle function, loss of physicalcapacity, loss of physical performance, impaired mobility, frailty,surgery, disability, risk of falling and risk of fall-related fracturesin a mammal. Preferably, said adult mammal is an elderly human.

The term “nutritional management” herein is to be understood as theprovision to a person of an amount of a nutrient or nutrients whichcorresponds to the recommended daily nutritional intake of that nutrientor those nutrients for that person, in particular an elderly person. Inorder to determine for an individual what the recommended amount of theliquid or semi-solid acid enteral nutritional composition according tothe invention to be taken should be to nutritionally manage this personin a desired manner, the skilled person has several detailed sources ofinformation at his disposal to achieve this. For instance, specifictables or other sources of information from governmental authorities,such as the Food and Nutrition Information Center of the United StatesDepartment of Agriculture, can be consulted to allow a person to beprovided with an amount of a nutrient or nutrients which corresponds tothe recommended daily nutritional intake of that nutrient or thosenutrients for that person, in particular an elderly person.

Dosage

In a specific embodiment, the nutritional composition according to theinvention has the form of a complete food, i.e. it can meet allnutritional needs of the user. As such, it preferably contains 1200 to2500 kcal per daily dosage. The daily dosage amounts are given withrespect to a daily energy supply of 2000 kcal to a healthy adult havinga body weight of 70 kg. For persons of different condition and differentbody weight, the levels should be adapted accordingly. It is understoodthat the average daily energy intake preferably is about 2000 kcal. Thenutritional composition, which may be a complete food, can be in theform of multiple dosage units, e.g. from 4 (e.g. 250 ml/unit) to 40(e.g. 20 ml/unit) per day for an energy supply of 2000 kcal/day usingthe liquid enteral nutritional composition according to the invention.

The liquid enteral nutritional composition can also be a foodsupplement, for example to be used in addition to a non-medical food.Preferably as a supplement, the liquid enteral nutritional compositioncontains per daily dosage less than 1500 kcal; in particular as asupplement, the liquid enteral nutritional composition contains 400 to1000 kcal per daily dose. The food supplement can be in the form ofmultiple dosage units, e.g. from 2 (250 ml/unit) to 10 (50 ml/unit) perday for an energy supply of 1000 kcal/day using the liquid enteralnutritional composition according to the invention.

In a further embodiment of the present invention, a unit dosagecomprises any amount of the liquid enteral nutritional compositionaccording to the invention between 10 ml and 250 ml, the end values ofthis range included, preferably any amount between 25 ml and 200 ml, theend values of this range included, more preferably any amount between 50ml and 150 ml, the end values of this range included, most preferablyabout 125 ml. For example, a person receiving 50 ml unit dosages can begiven 10 unit dosages per day to provide nutritional support using theliquid enteral nutritional composition according to the invention.Alternatively a person receiving 125 ml unit dosages can be given 4 or 5or 6 or 7 or 8 unit dosages per day to provide nutritional support usingthe liquid enteral nutritional composition according to the invention.Such small dosage units are preferred because of better compliance.

In a further embodiment, the nutritional composition is administered as1 to 2 servings daily, each serving comprising between 80 and 200 kcal,preferably about 125 kcal, preferably about 150 kcal. Preferably, thenutritional composition is administered as one serving daily. Using anutritional composition in a liquid or spoonable form, the serving maycomprise 30 to 250 ml of nutritional composition according to theinvention, most preferably 200 ml per serving.

In a further embodiment of the present invention, the composition isprovided in a ready to use liquid form and does not requirereconstitution or mixing prior to use. The composition according to theinvention can be tube fed or administered orally. For example, thecomposition according to the invention can be provided in a can, onspike, and hang bag.

In a further embodiment of the present invention, the compositionaccording to the invention is packaged. The packaging may have anysuitable form, for example a block-shaped carton, e.g. to be emptiedwith a straw; a carton or plastic beaker with removable cover; asmall-sized bottle for example for the 80 ml to 200 ml range, and smallcups for example for the 10 ml to 30 ml range. Another suitablepackaging mode is inclusion of small volumes of liquid (e.g. 10 ml to 20ml) in edible solid or semi-solid hulls or capsules, for examplegelatine-like coverings and the like.

The invention will now be described by way of examples; these are notmeant to be limiting.

EXPERIMENTAL

Example 1: According to the inventionExample 2: According to the inventionExample 3: According to Example 1 with about 130 mg/100 ml divalentmetal cationsExample 4: According to Example 1 with 100 weight % non-acidified WPI.Example 5: According to Example 1 with 40 weight % acidified and 60weight % non-acidified WPIExample 6: According to Example 1 with 116 g/L whey and 400 mg/100 ml CaExample 7: According to Example 1 with 160 g/L whey and 400 mg/100 ml CaExample 8: According to Example 1 with higher amounts of fat.Example 9: According to Example 1 with pH=3.7 during DSI.Example 10: According to Example 1 with pH=4.9 during DSI.Example 11: (using a reference method, comparative to Example 1 but withhomogenisation after DSI treatmentExample 12: (using a reference method, comparative to Example 1 butwithout DSI treatment)Example 13: (Comparative, as Example 1 except for a divalent cationsconcentration of less than 100 mg/100 ml)Example 14: according to the invention (spoonable product)Example 15: according to the invention (spoonable product):

Example 1

To obtain 20 L final product 909 g WPI and 1412 g pre-acidified WPI weredissolved to dissolve a total amount of 2000 g whey protein in 12.7 kgdemineralized water. This mixture was mixed under gentle stirring toavoid excessive foam formation. In this mixture also 8 g carotenoidmixture, 79 g L-leucine, 6 g L-isoleucine, 23 g L-valine, 280 g GOSsyrup (45% w/w pure), 14 g Inulin fibre source (97% w/w pure), 31 g lowviscosity pectin source (90% w/w pure), 60 g high methoxy pectin source(35% w/w pure) and 505 g sucrose were dissolved. The mixture was stirredfor about 2 hours at ambient temperature until all macro nutrients weredissolved or evenly dispersed. 91 g calcium hydroxide was added to 1346g demineralized water and stirred with a stirring rod for 5 minutes. 173g citric acid monohydrate was added to the calcium hydroxide solutionand after that the mixture is stirred again for 5 minutes at ambienttemperature. 7 g choline chloride, 6 g sodium ascorbate, 9 g potassiumchloride, 11 g tri-potassium citrate 1 aq, 30 g magnesium chloride 2 aqand 34 g tri-sodium citrate 2 aq were added to 492 g demineralized waterand stirred for about 1 hour at ambient temperature until all mineralswere dissolved or evenly dispersed. The solution containing the macroingredients was mixed with the solution containing calcium hydroxide andwith the solution containing the other minerals. This combined solutionwas stirred for several minutes to ensure evenly distribution of allcomponents in the solution. The pH of this solution was set at pH 4.3using lactic acid quantum satis. The pH was measured at ambienttemperature with an electrode directly in the solution. 19 g soylecithin and 278 g rapeseed oil were mixed at ambient temperature andsubsequently heated until 60° C. in a water bath. The oil mixture wasadded to the previous solution. A pre-emulsion was formed by mixing theoil trough the solution using an ultra thurrax. The newly formed mixturewas homogenized at 60° C. with 550+50 bar and cooled to ambienttemperature. The emulsion was preheated until 60° C., heated using DSIto 115° C. for 4 seconds and flash cooled to 60° C. The emulsion wascooled until ambient temperature. After this treatment the pH of theemulsion was adjusted to 4.0 using lactic acid quantum satis. The pH wasmeasured at ambient temperature with an electrode directly in thesolution. The dry matter of the product was adjusted by addingdemineralized water to obtain the required final dry matter. Theemulsion was mixed for about 5 minutes to evenly distribute the waterand the emulsion. The emulsion was pre-heated to 60° C. using a plateheat exchanger and given an UHT treatment. Product was filled in bottlesin a sterile cabinet.

Example 2

To obtain 20 L final product 909 g WPI and 1412 g pre-acidified WPI weredissolved to dissolve a total amount of 2000 g whey protein in 11.7 kgtap water. This mixture was mixed under gentle stirring to avoidexcessive foam formation. In this mixture also 79 g L-leucine, 6 gL-isoleucine, 23 g L-valine, 280 g GOS syrup (45% w/w pure) and 14 gInulin fibre source (97% w/w pure) were dissolved. The mixture wasstirred for about 2 hours at ambient temperature until all macronutrients were dissolved or evenly dispersed. 8 g carotenoid mixture wasdissolved in 75 g tap water, stirred for 5 minutes at ambienttemperature and added to the macro nutrient mixture. 85 g calciumhydroxide was added to 1255 g tap water and stirred with a stirring rodfor 1 minute. 161 g citric acid monohydrate was added to the calciumhydroxide solution and after that the mixture is stirred again for 3minutes at ambient temperature. 7 g choline chloride, 8 g calciumchloride 2 aq, 24 g tri-potassium citrate 1 aq, 29 g magnesium chloride2 aq and 30 g tri-sodium citrate 2 aq were added to 490 g tap water andstirred for about 1 hour at ambient temperature until all minerals weredissolved or evenly dispersed. 5 g mineral premix and 5 g tri-sodiumcitrate 2 aq were added to 90 g tap water and stirred for about 1 hourat ambient temperature until all minerals were dissolved or evenlydispersed. The solution containing the macro ingredients was mixed withthe solution containing calcium hydroxide and with the other two mineralcontaining solutions. This combined solution was stirred for severalminutes to ensure evenly distribution of all components in the solution.45 g low viscosity pectin source (90% w/w pure), 80 g high methoxypectin source (35% w/w pure) and 430 g sucrose are dry blended and addedto the above described solution under gentle stirring at ambienttemperature. The pH of this solution was set at pH 4.3 using lactic acidquantum satis. The pH was measured at ambient temperature with anelectrode directly in the solution. 19 g soy lecithin and 278 g rapeseedoil were mixed at ambient temperature and subsequently heated until 60°C. in a water bath. The oil mixture was added to the previous solution.A pre-emulsion was formed by mixing the oil trough the solution using anultra thurrax. The newly formed mixture was homogenized at 60° C. with550+50 bar. The emulsion was preheated until 60° C., heated using DSI to110° C. for 4 seconds and flash cooled to 60° C. The emulsion was cooleduntil ambient temperature. After this treatment the pH of the emulsionwas adjusted to 4.0 using lactic acid quantum satis. The pH was measuredat ambient temperature with an electrode directly in the solution. Thedry matter of the product was adjusted by adding tap water to obtain therequired final dry matter. The emulsion was mixed for about 5 minutes toevenly distribute the water and the emulsion. 5 g sodium ascorbate wasadded to the emulsion under gentle stirring. The emulsion was pre-heatedto 60° C. using a tube heat exchanger and given an UHT treatment.Product was filled in bottles in a sterile cabinet.

Example 3 Example 1 with about 130 mg/100 ml Divalent Metal Cations

To obtain 20 L final product 909 g WPI and 1412 g pre-acidified WPI weredissolved to dissolve a total amount of 2000 g whey protein in 12.7 kgdemineralized water. Before addition of the proteins, 8 g caroteinoidmixture was added to the water. This mixture was mixed under gentlestirring to avoid excessive foam formation. In this mixture also 79 gL-leucine, 6 g L-isoleucine, 23 g L-valine, 280 g GOS syrup (45% w/wpure), 15 g Inulin fibre source (97% w/w pure), 31 g low viscositypectin source (90% w/w pure), 60 g high methoxy pectin source (35% w/wpure) and 505 g sucrose were dissolved. The mixture was stirred forabout 2 hours at ambient temperature until all macro nutrients weredissolved or evenly dispersed. 46 g calcium hydroxide was added to 673 gdemineralized water and stirred with a stirring rod for 5 minutes. 86 gcitric acid monohydrate was added to the calcium hydroxide solution andafter that the mixture is stirred again for 5 minutes at ambienttemperature. 7 g choline chloride, 6 g sodium ascorbate, 9 g potassiumchloride, 6 g tri-potassium citrate 1 aq, 15 g magnesium chloride 2 aqand 17 g tri-sodium citrate 2 aq were added to 256 g demineralized waterand stirred for about 1 hour at ambient temperature until all mineralswere dissolved or evenly dispersed. The solution containing the macroingredients was mixed with the solution containing calcium hydroxide andwith the solution containing the other minerals. This combined solutionwas stirred for several minutes to ensure evenly distribution of allcomponents in the solution. The pH of this solution was set at pH 4.3using lactic acid quantum satis. The pH was measured at ambienttemperature with an electrode directly in the solution. 19 g soylecithin and 278 g rapeseed oil were mixed at ambient temperature andsubsequently heated until 60° C. in a water bath. The oil mixture wasadded to the previous solution. A pre-emulsion was formed by mixing theoil trough the solution using an ultra thurrax. The newly formed mixturewas homogenized at 60° C. with 550+50 bar and cooled to ambienttemperature. The emulsion was preheated until 60° C., heated using DSIto 115° C. for 4 seconds and flash cooled to 60° C. The emulsion wascooled until ambient temperature. After this treatment the pH of theemulsion was adjusted to 4.0 using lactic acid quantum satis. The pH wasmeasured at ambient temperature with an electrode directly in thesolution. The dry matter of the product was adjusted by addingdemineralized water to obtain the required final dry matter. Theemulsion was mixed for about 5 minutes to evenly distribute the waterand the emulsion. The emulsion was pre-heated to 60° C. using a plateheat exchanger and given an UHT treatment. Product was filled in bottlesin a sterile cabinet.

Example 4 Example 1 with 100 Weight % Non-Acidified WPI

To obtain 20 L final product 2273 g WPI was dissolved to dissolve atotal amount of 2000 g whey protein in 12.7 kg demineralized water. Thismixture was mixed under gentle stirring to avoid excessive foamformation. In this mixture also 79 g L-leucine, 6 g L-isoleucine, 23 gL-valine, 280 g GOS syrup (45% w/w pure), 15 g Inulin fibre source (97%w/w pure), 31 g low viscosity pectin source (90% w/w pure), 80 g highmethoxy pectin source (35% w/w pure) and 505 g sucrose were dissolved.The mixture was stirred for about 2 hours at ambient temperature untilall macro nutrients were dissolved or evenly dispersed. 49 g calciumhydroxide was added to 715 g demineralized water and stirred with astirring rod for 5 minutes. 92 g citric acid monohydrate was added tothe calcium hydroxide solution and after that the mixture is stirredagain for 5 minutes at ambient temperature. 7 g choline chloride, 6 gsodium ascorbate, 11 g tri-sodium citrate 2 aq, 26 g magnesium hydrogenphosphate 3 aq, 35 g penta calcium triphosphate and 32 g calciumchloride were added to 583 g demineralized water and stirred for about 1hour at ambient temperature until all minerals were dissolved or evenlydispersed. The solution containing the macro ingredients was mixed withthe solution containing calcium hydroxide and with the solutioncontaining the other minerals. This combined solution was stirred forseveral minutes to ensure evenly distribution of all components in thesolution. The pH of this solution was set at pH 4.9 using lactic acidquantum satis. The pH was measured at ambient temperature with anelectrode directly in the solution. 19 g soy lecithin and 278 g rapeseedoil were mixed at ambient temperature and subsequently heated until 60°C. in a water bath. The oil mixture was added to the previous solution.A pre-emulsion was formed by mixing the oil trough the solution using anultra thurrax. The newly formed mixture was homogenized at 60° C. with550+50 bar and cooled to ambient temperature. The emulsion was preheateduntil 60° C., heated using DSI to 110° C. for 4 seconds and flash cooledto 60° C. The emulsion was cooled until ambient temperature. After thistreatment the pH of the emulsion was adjusted to 4.0 using lactic acidquantum satis. The pH was measured at ambient temperature with anelectrode directly in the solution. The dry matter of the product wasadjusted by adding demineralized water to obtain the required final drymatter. The emulsion was mixed for about 5 minutes to evenly distributethe water and the emulsion. The emulsion was pre-heated to 60° C. usinga plate heat exchanger and given an UHT treatment. Product was filled inbottles in a sterile cabinet.

Example 5 Example 1 with 40 Weight % Acidified and 60 Weight %Non-Acidified WPI

To obtain 20 L final product 1364 g WPI and 941 g pre-acidified WPI weredissolved to dissolve a total amount of 2000 g whey protein in 12.6 kgdemineralized water. This mixture was mixed under gentle stirring toavoid excessive foam formation. In this mixture also 79 g L-leucine, 6 gL-isoleucine, 23 g L-valine, 280 g GOS syrup (45% w/w pure), 15 g Inulinfibre source (97% w/w pure), 31 g low viscosity pectin source (90% w/wpure), 80 g high methoxy pectin source (35% w/w pure) and 505 g sucrosewere dissolved. The mixture was stirred for about 2 hours at ambienttemperature until all macro nutrients were dissolved or evenlydispersed. 86 g calcium hydroxide was added to 1265 g demineralizedwater and stirred with a stirring rod for 5 minutes. 162 g citric acidmonohydrate was added to the calcium hydroxide solution and after thatthe mixture is stirred again for 5 minutes at ambient temperature. 7 gcholine chloride, 6 g sodium ascorbate, 9 g tri potassium citrate laq,30 g magnesium chloride 6 aq, 25 g tri sodium citrate and 25 g calciumchloride were added to 438 g demineralized water and stirred for about 1hour at ambient temperature until all minerals were dissolved or evenlydispersed. The solution containing the macro ingredients was mixed withthe solution containing calcium hydroxide and with the solutioncontaining the other minerals. This combined solution was stirred forseveral minutes to ensure evenly distribution of all components in thesolution. The pH of this solution was set at pH 4.9 using lactic acidquantum satis. The pH was measured at ambient temperature with anelectrode directly in the solution. 19 g soy lecithin and 278 g rapeseedoil were mixed at ambient temperature and subsequently heated until 60°C. in a water bath. The oil mixture was added to the previous solution.A pre-emulsion was formed by mixing the oil trough the solution using anultra thurrax. The newly formed mixture was homogenized at 60° C. with550+50 bar and cooled to ambient temperature. The emulsion was preheateduntil 60° C., heated using DSI to 110° C. for 4 seconds and flash cooledto 60° C. The emulsion was cooled until ambient temperature. After thistreatment the pH of the emulsion was adjusted to 4.0 using lactic acidquantum satis. The pH was measured at ambient temperature with anelectrode directly in the solution. The dry matter of the product wasadjusted by adding demineralized water to obtain the required final drymatter. The emulsion was mixed for about 5 minutes to evenly distributethe water and the emulsion. The emulsion was pre-heated to 60° C. usinga plate heat exchanger and given an UHT treatment. Product was filled inbottles in a sterile cabinet.

Example 6 Example 1 with 116 g/L Whey and 400 mg/100 ml Ca

To obtain 20 L final product 1055 g WPI and 1638 g pre-acidified WPIwere dissolved to dissolve a total amount of 2320 g whey protein in 12.1kg demineralized water. This mixture was mixed under gentle stirring toavoid excessive foam formation. In this mixture also 8 g carotenoidmixture, 216 g L-leucine, 444 g GOS syrup (45% w/w pure), 23 g Inulinfibre source (97% w/w pure), 49 g low viscosity pectin source (90% w/wpure), 60 g high methoxy pectin source (35% w/w pure) and 1088 g sucrosewere dissolved. The mixture was stirred for about 2 hours at ambienttemperature until all macro nutrients were dissolved or evenlydispersed. 149 g calcium hydroxide was added to 2191 g demineralizedwater and stirred with a stirring rod for 5 minutes. 281 g citric acidmonohydrate was added to the calcium hydroxide solution and after thatthe mixture is stirred again for 5 minutes at ambient temperature. 12 gcholine chloride, 9 g sodium ascorbate, 42 g magnesium hydrogenphosphate 2 aq, 33 g sodium chloride and 33 g dipotassiumhydrogenphosphate were added to 641 g demineralized water and stirredfor about 1 hour at ambient temperature until all minerals weredissolved or evenly dispersed. The solution containing the macroingredients was mixed with the solution containing calcium hydroxide andwith the solution containing the other minerals. This combined solutionwas stirred for several minutes to ensure evenly distribution of allcomponents in the solution. The pH of this solution was set at pH 4.3using lactic acid quantum satis. The pH was measured at ambienttemperature with an electrode directly in the solution. 25 g soylecithin and 372 g rapeseed oil were mixed at ambient temperature andsubsequently heated until 60° C. in a water bath. The oil mixture wasadded to the previous solution. A pre-emulsion was formed by mixing theoil trough the solution using an ultra thurrax. The newly formed mixturewas homogenized at 60° C. with 550+50 bar and cooled to ambienttemperature. The emulsion was preheated until 60° C., heated using DSIto 115° C. for 4 seconds and flash cooled to 60° C. The emulsion wascooled until ambient temperature. After this treatment the pH of theemulsion was adjusted to 4.0 using lactic acid quantum satis. The pH wasmeasured at ambient temperature with an electrode directly in thesolution. The dry matter of the product was adjusted by addingdemineralized water to obtain the required final dry matter. Theemulsion was mixed for about 5 minutes to evenly distribute the waterand the emulsion. The emulsion was pre-heated to 60° C. using a plateheat exchanger and given an UHT treatment. Product was filled in bottlesin a sterile cabinet.

Example 7 Example 1 with 160 g/L Whey and 400 mg/100 ml Ca

To obtain 20 L final product 3546 g WPI was dissolved to dissolve atotal amount of 3200 g whey protein in 13.0 kg demineralized water. Thismixture was mixed under gentle stirring to avoid excessive foamformation. In this mixture also 80 g L-leucine, 18 g L-isoleucine, 10 gL-valine, 444 g GOS syrup (45% w/w pure) and 23 g Inulin fibre source(97% w/w pure) were dissolved. After all ingredients were dissolved orevenly dispersed the following ingredients were dissolved in thismixture: 51 g low viscosity pectin source (90% w/w pure), 80 g highmethoxy pectin source (35% w/w pure) and 963 g sucrose. The mixture wasstirred for about 2 hours at ambient temperature until all macronutrients were dissolved or evenly dispersed. 82 g calcium hydroxide wasadded to 1210 g demineralized water and stirred with a stirring rod for5 minutes. 155 g citric acid monohydrate was added to the calciumhydroxide solution and after that the mixture is stirred again for 5minutes at ambient temperature. 12 g choline chloride, 9 g sodiumascorbate, 36 g magnesium hydrogen phosphate 2 aq, 42 g penta calciumtriphosphate, 5 g potassium chloride and 10 g tri-potassium citrate 1 aqwere added to 825 g demineralized water and stirred for about 1 hour atambient temperature until all minerals were dissolved or evenlydispersed. The solution containing the macro ingredients was mixed withthe solution containing calcium hydroxide and with the solutioncontaining the other minerals. This combined solution was stirred forseveral minutes to ensure evenly distribution of all components in thesolution. The pH of this solution was set at pH 4.3 using lactic acidquantum satis. The pH was measured at ambient temperature with anelectrode directly in the solution. 30 g soy lecithin and 442 g rapeseedoil were mixed at ambient temperature and subsequently heated until 60°C. in a water bath. The oil mixture was added to the previous solution.A pre-emulsion was formed by mixing the oil trough the solution using anultra thurrax. The newly formed mixture was homogenized at 20° C. with550+50 bar and cooled to ambient temperature. The emulsion was heatedusing DSI to 115° C. for 4 seconds and flash cooled to 60° C. Theemulsion was cooled until ambient temperature. After this treatment thepH of the emulsion was adjusted to 4.0 using lactic acid quantum satis.The pH was measured at ambient temperature with an electrode directly inthe solution. The dry matter of the product was adjusted by addingdemineralized water to obtain the required final dry matter. Theemulsion was mixed for about 5 minutes to evenly distribute the waterand the emulsion. The emulsion was pre-heated to 60° C. using a plateheat exchanger and given an UHT treatment. Product was filled in bottlesin a sterile cabinet.

Example 8 Example 1 with Higher Amounts of Fat

To obtain 20 L final product 909 g WPI and 1412 g pre-acidified WPI weredissolved to dissolve a total amount of 2000 g whey protein in 12.7 kgdemineralized water. This mixture was mixed under gentle stirring toavoid excessive foam formation. In this mixture also 8 g carotenoidmixture, 79 g L-leucine, 6 g L-isoleucine, 23 g L-valine, 280 g GOSsyrup (45% w/w pure), 14 g Inulin fibre source (97% w/w pure), 31 g lowviscosity pectin source (90% w/w pure), 60 g high methoxy pectin source(35% w/w pure) and 2738 g sucrose were dissolved. The mixture wasstirred for about 2 hours at ambient temperature until all macronutrients were dissolved or evenly dispersed. 91 g calcium hydroxide wasadded to 1346 g demineralized water and stirred with a stirring rod for5 minutes. 173 g citric acid monohydrate was added to the calciumhydroxide solution and after that the mixture is stirred again for 5minutes at ambient temperature. 7 g choline chloride, 6 g sodiumascorbate, 9 g potassium chloride, 11 g tri-potassium citrate 1 aq, 30 gmagnesium chloride 2 aq and 34 g tri-sodium citrate 2 aq were added to492 g demineralized water and stirred for about 1 hour at ambienttemperature until all minerals were dissolved or evenly dispersed. Thesolution containing the macro ingredients was mixed with the solutioncontaining calcium hydroxide and with the solution containing the otherminerals. This combined solution was stirred for several minutes toensure evenly distribution of all components in the solution. The pH ofthis solution was set at pH 4.3 using lactic acid quantum satis. The pHwas measured at ambient temperature with an electrode directly in thesolution. 80 g soy lecithin and 1120 g rapeseed oil were mixed atambient temperature and subsequently heated until 60° C. in a waterbath. The oil mixture was added to the previous solution. A pre-emulsionwas formed by mixing the oil trough the solution using an ultra thurrax.The newly formed mixture was homogenized at 60° C. with 550+50 bar andcooled to ambient temperature. The emulsion was pre-heated until 60° C.,heated using DSI to 115° C. for 4 seconds and flash cooled to 60° C. Theemulsion was cooled until ambient temperature. After this treatment thepH of the emulsion was adjusted to 4.0 using lactic acid quantum satis.The pH was measured at ambient temperature with an electrode directly inthe solution. The dry matter of the product was adjusted by addingdemineralized water to obtain the required final dry matter. Theemulsion was mixed for about 5 minutes to evenly distribute the waterand the emulsion. The emulsion was pre-heated to 60° C. using a plateheat exchanger and given an UHT treatment. Product was filled in bottlesin a sterile cabinet.

Example 9 Example 1 with pH=3.7 During DSI

To obtain 20 L final product 909 g WPI and 1412 g pre-acidified WPI weredissolved to dissolve a total amount of 2000 g whey protein in 13.5 kgdemineralized water. This mixture was mixed under gentle stirring toavoid excessive foam formation. In this mixture also 8 g carotenoidmixture, 79 g L-leucine, 6 g L-isoleucine, 23 g L-valine, 280 g GOSsyrup (45% w/w pure), 14 g Inulin fibre source (97% w/w pure), 31 g lowviscosity pectin source (90% w/w pure), 60 g high methoxy pectin source(35% w/w pure) and 505 g sucrose were dissolved. The mixture was stirredfor about 2 hours at ambient temperature until all macro nutrients weredissolved or evenly dispersed. 91 g calcium hydroxide was added to 1346g demineralized water and stirred with a stirring rod for 5 minutes. 173g citric acid monohydrate was added to the calcium hydroxide solutionand after that the mixture is stirred again for 5 minutes at ambienttemperature. 7 g choline chloride, 6 g sodium ascorbate, 9 g potassiumchloride, 11 g tri-potassium citrate 1 aq, 30 g magnesium chloride 2 aqand 34 g tri-sodium citrate 2 aq were added to 492 g demineralized waterand stirred for about 1 hour at ambient temperature until all mineralswere dissolved or evenly dispersed. The solution containing the macroingredients was mixed with the solution containing calcium hydroxide andwith the solution containing the other minerals. This combined solutionwas stirred for several minutes to ensure evenly distribution of allcomponents in the solution. The pH of this solution was set at pH 3.7using lactic acid quantum satis. The pH was measured at ambienttemperature with an electrode directly in the solution. 19 g soylecithin and 278 g rapeseed oil were mixed at ambient temperature andsubsequently heated until 60° C. in a water bath. The oil mixture wasadded to the previous solution. A pre-emulsion was formed by mixing theoil trough the solution using an ultra thurrax. The newly formed mixturewas homogenized at 60° C. with 550+50 bar and cooled to ambienttemperature. The emulsion was preheated until 60° C., heated using DSIto 115° C. for 4 seconds and flash cooled to 60° C. The emulsion wascooled until ambient temperature. After this treatment the pH of theemulsion was adjusted to 4.0 using a potassium hydroxide solutionquantum satis. The pH was measured at ambient temperature with anelectrode directly in the solution. The dry matter of the product wasadjusted by adding demineralized water to obtain the required final drymatter. The emulsion was mixed for about 5 minutes to evenly distributethe water and the emulsion. The emulsion was pre-heated to 60° C. usinga plate heat exchanger and given an UHT treatment. Product was filled inbottles in a sterile cabinet.

Example 10 Example 1 with pH=4.9 During DSI

To obtain 20 L final product 909 g WPI and 1412 g pre-acidified WPI weredissolved to dissolve a total amount of 2000 g whey protein in 13.5 kgdemineralized water. This mixture was mixed under gentle stirring toavoid excessive foam formation. In this mixture also 8 g carotenoidmixture, 79 g L-leucine, 6 g L-isoleucine, 23 g L-valine, 280 g GOSsyrup (45% w/w pure), 14 g Inulin fibre source (97% w/w pure), 31 g lowviscosity pectin source (90% w/w pure), 60 g high methoxy pectin source(35% w/w pure) and 505 g sucrose were dissolved. The mixture was stirredfor about 2 hours at ambient temperature until all macro nutrients weredissolved or evenly dispersed. 91 g calcium hydroxide was added to 1346g demineralized water and stirred with a stirring rod for 5 minutes. 173g citric acid monohydrate was added to the calcium hydroxide solutionand after that the mixture is stirred again for 5 minutes at ambienttemperature. 7 g choline chloride, 6 g sodium ascorbate, 9 g potassiumchloride, 11 g tri-potassium citrate 1 aq, 30 g magnesium chloride 2 aqand 34 g tri-sodium citrate 2 aq were added to 492 g demineralized waterand stirred for about 1 hour at ambient temperature until all mineralswere dissolved or evenly dispersed. The solution containing the macroingredients was mixed with the solution containing calcium hydroxide andwith the solution containing the other minerals. This combined solutionwas stirred for several minutes to ensure evenly distribution of allcomponents in the solution. The pH of this solution was set at pH 4.9using a potassium hydroxide solution quantum satis. The pH was measuredat ambient temperature with an electrode directly in the solution. 19 gsoy lecithin and 278 g rapeseed oil were mixed at ambient temperatureand subsequently heated until 60° C. in a water bath. The oil mixturewas added to the previous solution. A pre-emulsion was formed by mixingthe oil trough the solution using an ultra thurrax. The newly formedmixture was homogenized at 60° C. with 550+50 bar and cooled to ambienttemperature. The emulsion was preheated until 60° C., heated using DSIto 115° C. for 4 seconds and flash cooled to 60° C. The emulsion wascooled until ambient temperature. After this treatment the pH of theemulsion was adjusted to 4.0 using lactic acid quantum satis. The pH wasmeasured at ambient temperature with an electrode directly in thesolution. The dry matter of the product was adjusted by addingdemineralized water to obtain the required final dry matter. Theemulsion was mixed for about 5 minutes to evenly distribute the waterand the emulsion. The emulsion was pre-heated to 60° C. using a plateheat exchanger and given an UHT treatment. Product was filled in bottlesin a sterile cabinet.

The results are summarized in Table 1.

TABLE 1 Summary of experiments Example 1 2 3 4 5 6 7 8 9 10 Experiment79.1 ZM36.1 78.2 59.1 59.3 1.4 72.1 81.5 106.1 106.5 code Whey protein10 10 10 10 10 11.6 16 10 10 10 (g/100 ml) Ratio 60:40 60:40 60:40 100:040:60 60:40 100:0 60:40 60:40 60:40 acidified: neutral WPI Calcium 250250 131 250 250 401 320 250 250 250 (mg/100 ml) Fat (g/100 ml) 1.5 1.51.5 1.5 1.5 2.0 2.4 6.0 1.5 1.5 Carbohydrates 4.8 4.7 4.3 5.2 5.6 8.38.7 15.9 4.8 4.8 (g/100 ml) Kcal/100 mL 75 75 74 75 77 103 120 160 75 75Viscosity 38 31 36 40 39 57 47 189 25 19 (mPa.s) pH during 4 4 4 4 4 4 44 3.7 4.9 DSI Astringency ++ ++ 0 ++ ++ ++ ++ ++ ++ ++ Sandiness ++ ++++ 0 ++ ++ 0 ++ ++ ++ Viscosity is measured at 20° C. at a shear rate of100 s-1. Astringency: low in astringency (++), astringent (0), veryastringent (−−) Sandiness: low in sandiness (++), sandy (0), very sandy(−−)

Example 11 As Example 1 with Homogenization AFTER DSI Treatment

To obtain 20 L final product 909 g WPI and 1412 g pre-acidified WPI weredissolved to dissolve a total amount of 2000 g whey protein in 11.7 kgtap water. This mixture was mixed under gentle stirring to avoidexcessive foam formation. In this mixture also 79 g L-leucine, 6 gL-isoleucine, 23 g L-valine, 280 g GOS syrup (45% w/w pure) and 14 gInulin fibre source (97% w/w pure) were dissolved. The mixture wasstirred for about 2 hours at ambient temperature until all macronutrients were dissolved or evenly dispersed. 8 g carotenoid mixture wasdissolved in 75 g tap water, stirred for 5 minutes at ambienttemperature and added to the macro nutrient mixture. 85 g calciumhydroxide was added to 1255 g tap water and stirred with a stirring rodfor 1 minute. 161 g citric acid monohydrate was added to the calciumhydroxide solution and after that the mixture is stirred again for 3minutes at ambient temperature. 7 g choline chloride, 8 g calciumchloride 2 aq, 24 g tri-potassium citrate 1 aq, 29 g magnesium chloride2 aq and 30 g tri-sodium citrate 2 aq were added to 490 g tap water andstirred for about 1 hour at ambient temperature until all minerals weredissolved or evenly dispersed. 5 g mineral premix and 5 g tri-sodiumcitrate 2 aq were added to 90 g tap water and stirred for about 1 hourat ambient temperature until all minerals were dissolved or evenlydispersed. The solution containing the macro ingredients was mixed withthe solution containing calcium hydroxide and with the other two mineralcontaining solutions. This combined solution was stirred for severalminutes to ensure evenly distribution of all components in the solution.45 g low viscosity pectin source (90% w/w pure), 80 g high methoxypectin source (35% w/w pure) and 430 g sucrose are dry blended and addedto the above described solution under gentle stirring at ambienttemperature. The pH of this solution was set at pH 4.3 using lactic acidquantum satis. The pH was measured at ambient temperature with anelectrode directly in the solution. 19 g soy lecithin and 278 g rapeseedoil were mixed at ambient temperature and subsequently heated until 60°C. in a water bath. The oil mixture was added to the previous solution.A pre-emulsion was formed by mixing the oil trough the solution using anultra thurrax. The newly formed mixture was preheated until 60° C.,heated using DSI to 110° C. for 4 seconds, flash cooled to 60° C. andhomogenized at 60° C. with 550+50 bar. The emulsion was cooled untilambient temperature. After this heat treatment the pH of the emulsionwas adjusted to 4.0 using lactic acid quantum satis. The pH was measuredat ambient temperature with an electrode directly in the solution. Thedry matter of the product was adjusted by adding tap water to obtain therequired final dry matter. The emulsion was mixed for about 5 minutes toevenly distribute the water and the emulsion. 5 g sodium ascorbate wasadded to the emulsion under gentle stirring. The emulsion was pre-heatedto 60° C. using a tube heat exchanger and given an UHT treatment.Product was filled in bottles in a sterile cabinet.

Example 12 As Example 1 without DSI Treatment

To obtain 20 L final product 909 g WPI and 1412 g pre-acidified WPI weredissolved to dissolve a total amount of 2000 g whey protein in 12.7 kgdemineralized water. This mixture was mixed under gentle stirring toavoid excessive foam formation. In this mixture also 8 g carotenoidmixture, 79 g L-leucine, 6 g L-isoleucine, 23 g L-valine, 280 g GOSsyrup (45% w/w pure), 14 g Inulin fibre source (97% w/w pure), 31 g lowviscosity pectin source (90% w/w pure), 60 g high methoxy pectin source(35% w/w pure) and 505 g sucrose were dissolved. The mixture was stirredfor about 2 hours at ambient temperature until all macro nutrients weredissolved or evenly dispersed. 91 g calcium hydroxide was added to 1346g demineralized water and stirred with a stirring rod for 5 minutes. 173g citric acid monohydrate was added to the calcium hydroxide solutionand after that the mixture is stirred again for 5 minutes at ambienttemperature. 7 g choline chloride, 6 g sodium ascorbate, 9 g potassiumchloride, 11 g tri-potassium citrate 1 aq, 30 g magnesium chloride 2 aqand 34 g tri-sodium citrate 2 aq were added to 492 g demineralized waterand stirred for about 1 hour at ambient temperature until all mineralswere dissolved or evenly dispersed. The solution containing the macroingredients was mixed with the solution containing calcium hydroxide andwith the solution containing the other minerals. This combined solutionwas stirred for several minutes to ensure evenly distribution of allcomponents in the solution. The pH of this solution was set at pH 4.3using lactic acid quantum satis. The pH was measured at ambienttemperature with an electrode directly in the solution. 19 g soylecithin and 278 g rapeseed oil were mixed at ambient temperature andsubsequently heated until 60° C. in a water bath. The oil mixture wasadded to the previous solution. A pre-emulsion was formed by mixing theoil trough the solution using an ultra thurrax. The newly formed mixturewas homogenized at 60° C. with 550+50 bar and cooled to ambienttemperature. The pH of the emulsion was adjusted to 4.0 using lacticacid quantum satis. The pH was measured at ambient temperature with anelectrode directly in the solution. The dry matter of the product wasadjusted by adding demineralized water to obtain the required final drymatter. The emulsion was mixed for about 5 minutes to evenly distributethe water and the emulsion. The emulsion was pre-heated to 60° C. usinga plate heat exchanger and given an UHT treatment. Product was filled inbottles in a sterile cabinet.

Example 13 Comparative, as Example 1, but with Less than 100 mg/100 mlDivalent Cations

To obtain 20 L final product 909 g WPI and 1412 g pre-acidified WPI weredissolved to dissolve a total amount of 2000 g whey protein in 12.7 kgdemineralized water. Before addition of the proteins, 8 g caroteinoidmixture was added to the water. This mixture was mixed under gentlestirring to avoid excessive foam formation. In this mixture also 79 gL-leucine, 6 g L-isoleucine, 23 g L-valine, 280 g GOS syrup (45% w/wpure), 15 g Inulin fibre source (97% w/w pure), 31 g low viscositypectin source (90% w/w pure), 60 g high methoxy pectin source (35% w/wpure) and 505 g sucrose were dissolved. The mixture was stirred forabout 2 hours at ambient temperature until all macro nutrients weredissolved or evenly dispersed. The pH of this solution was set at pH 4.3using lactic acid quantum satis. The pH was measured at ambienttemperature with an electrode directly in the solution. 19 g soylecithin and 278 g rapeseed oil were mixed at ambient temperature andsubsequently heated until 60° C. in a water bath. The oil mixture wasadded to the previous solution. A pre-emulsion was formed by mixing theoil trough the solution using an ultra thurrax. The newly formed mixturewas homogenized at 60° C. with 550+50 bar and cooled to ambienttemperature. The emulsion was preheated until 60° C., heated using DSIto 115° C. for 4 seconds and flash cooled to 60° C. The emulsion wascooled until ambient temperature. After this treatment the pH of theemulsion was adjusted to 4.0 using lactic acid quantum satis. The pH wasmeasured at ambient temperature with an electrode directly in thesolution. The dry matter of the product was adjusted by addingdemineralized water to obtain the required final dry matter. Theemulsion was mixed for about 5 minutes to evenly distribute the waterand the emulsion. The emulsion was pre-heated to 60° C. using a plateheat exchanger and given an UHT treatment. Product was filled in bottlesin a sterile cabinet. The results are summarized in Table 2.

TABLE 2 Summary of Examples 11-13 Ex 11 (ref. Ex12 (ref. Ex 13 Examplemethod) method) (comparative) Experiment code ZM36.2 79.2 78.1 Wheyprotein (g/100 ml) 10 10 10 Ratio acidified:neutral whey 60:40 60:4060:40 Calcium (mg/100 ml) 250 250 12 Fat (g/100 ml) 1.5 1.5 1.5Carbohydrates (g/100 ml) 4.4 4.8 3.8 Kcal/100 mL 75 75 72 Viscosity (mPa· s) 35 118 92 Astringency 0 −− −− pH during DSI 4 no DSI 4 Sandiness −−0 0 Viscosity is measured at 20° C. at a shear rate of 100 s−1.Astringency: low in astringency (++), astringent (0), very astringent(−−) Sandiness: low in sandiness (++), sandy (0), very sandy (−−)

Example 14 Preparation of a Spoonable Composition

To obtain 20 L final product 909 g WPI and 1412 g pre-acidified WPI,giving a total amount of 2000 g whey protein, were dissolved in 12.6 kgdemineralized water. This mixture was mixed under gentle stirring toavoid excessive foam formation. In this mixture also 8 g carotenoidmixture, 79 g L-leucine, 6 g L-isoleucine, 23 g L-valine, 280 g GOSsyrup (45% w/w pure), 14 g Inulin fibre source (97% w/w pure), 31 g lowviscosity pectin source (90% w/w pure), 200 g high methoxy pectin source(35% w/w pure) and 505 g sucrose were dissolved. The mixture was stirredfor about 2 hours at ambient temperature until all macro nutrients weredissolved or evenly dispersed. 91 g calcium hydroxide was added to 1346g demineralized water and stirred with a stirring rod for 5 minutes. 173g citric acid monohydrate was added to the calcium hydroxide solutionand after that the mixture is stirred again for 5 minutes at ambienttemperature. 7 g choline chloride, 6 g sodium ascorbate, 9 g potassiumchloride, 11 g tri-potassium citrate 1 aq, 30 g magnesium chloride 2 aqand 34 g tri-sodium citrate 2 aq were added to 492 g demineralized waterand stirred for about 1 hour at ambient temperature until all mineralswere dissolved or evenly dispersed. The solution containing the macroingredients was mixed with the solution containing calcium hydroxide andwith the solution containing the other minerals. This combined solutionwas stirred for several minutes to ensure evenly distribution of allcomponents in the solution. The pH of this solution was set at pH 4.0using lactic acid quantum satis. The pH was measured at ambienttemperature with an electrode directly in the solution. 19 g soylecithin and 278 g rapeseed oil were mixed at ambient temperature andsubsequently heated until 60° C. in a water bath. The oil mixture wasadded to the previous solution. A pre-emulsion was formed by mixing theoil trough the solution using an ultra thurrax. The newly formed mixturewas homogenized at 60° C. with 550+50 bar and cooled to ambienttemperature. The emulsion was preheated until 60° C., heated using DSIto 115° C. for 4 seconds and flash cooled to 60° C. The emulsion wascooled until ambient temperature. After this treatment the pH of theemulsion was checked and if needed adjusted to 4.0 using lactic acidquantum satis. The pH was measured at ambient temperature with anelectrode directly in the solution. The dry matter of the product wasadjusted by adding demineralized water to obtain the required final drymatter. The emulsion was mixed for about 5 minutes to evenly distributethe water and the emulsion. The emulsion was pre-heated to 60° C. usinga plate heat exchanger and given an UHT treatment. Product was filled inplastic cups in a sterile cabinet.

Example 15 Preparation of a Spoonable Composition

To obtain 20 L final product 909 g WPI and 1412 g pre-acidified WPI,giving a total amount of 2000 g whey protein, were dissolved in 12.5 kgdemineralized water. This mixture was mixed under gentle stirring toavoid excessive foam formation. In this mixture also 8 g carotenoidmixture, 79 g L-leucine, 6 g L-isoleucine, 23 g L-valine, 280 g GOSsyrup (45% w/w pure), 14 g Inulin fibre source (97% w/w pure), 31 g lowviscosity pectin source (90% w/w pure), 60 g high methoxy pectin source(35% w/w pure), 200 g modified starch (E1442) and 505 g sucrose weredissolved. The mixture was stirred for about 2 hours at ambienttemperature until all macro nutrients were dissolved or evenlydispersed. 91 g calcium hydroxide was added to 1346 g demineralizedwater and stirred with a stirring rod for 5 minutes. 173 g citric acidmonohydrate was added to the calcium hydroxide solution and after thatthe mixture is stirred again for 5 minutes at ambient temperature. 7 gcholine chloride, 6 g sodium ascorbate, 9 g potassium chloride, 11 gtri-potassium citrate laq, 30 g magnesium chloride 2 aq and 34 gtri-sodium citrate 2 aq were added to 492 g demineralized water andstirred for about 1 hour at ambient temperature until all minerals weredissolved or evenly dispersed. The solution containing the macroingredients was mixed with the solution containing calcium hydroxide andwith the solution containing the other minerals. This combined solutionwas stirred for several minutes to ensure evenly distribution of allcomponents in the solution. The pH of this solution was set at pH 4.0using lactic acid quantum satis. The pH was measured at ambienttemperature with an electrode directly in the solution. 19 g soylecithin and 278 g rapeseed oil were mixed at ambient temperature andsubsequently heated until 60° C. in a water bath. The oil mixture wasadded to the previous solution. A pre-emulsion was formed by mixing theoil trough the solution using an ultra thurrax. The newly formed mixturewas homogenized at 60° C. with 550+50 bar and cooled to ambienttemperature. The emulsion was preheated until 60° C., heated using DSIto 115° C. for 4 seconds and flash cooled to 60° C. The emulsion wascooled until ambient temperature. After this treatment the pH of theemulsion was checked and if needed adjusted to 4.0 using lactic acidquantum satis. The pH was measured at ambient temperature with anelectrode directly in the solution. The dry matter of the product wasadjusted by adding demineralized water to obtain the required final drymatter. The emulsion was mixed for about 5 minutes to evenly distributethe water and the emulsion. The emulsion was pre-heated to 60° C. usinga plate heat exchanger and given an UHT treatment. Product was filled inplastic cups in a sterile cabinet.

Example 14 15 Experiment code 6.1 6.2 Whey protein (g/100 ml) 10 10Ratio acidified:neutral WPI 40:60 40:60 Calcium (mg/100 ml) 250 250 Fat(g/ml) 1.5 1.5 Carbohydrates (g/ml) 5.1 5.8 Stabilizers 0.35% HM pectin0.1% HM pectin 1% Starch Kcal/100 mL 77 79 Viscosity 308 (0 wk) 246 (0wk) (mPa · s) 579 (6 wk) 490 (6 wk) (time) pH during DSI 4 4 Astringency++ ++ Sandiness ++ ++

2. Nutritional Compositions

The following nutritional composition according to the invention issuitable for the prevention or treatment of a disease in an elderlymammal, which involves muscle protein synthesis.

TABLE 3 Example of a liquid sip feed composition (200 ml Serving size)Liquid sip feed Liquid sip feed Ingredient (per 100 kcal) (per 100 ml)Energy (kcal) 100 75 protein (En %) 56 56 fat (En %) 18 18 digestiblecarb (En %) 23 23 indigestible carb (En %) 3 3 Total protein (g) 14.010.5 Intact whey protein (g) 13.3 (95 wt %)  10.0 (95 wt %)  (wt % ofproteinaceous matter) Free leucine (g) 0.5 (26 wt %) 0.4 (26 wt %) (wt %of total leucine) Total leucine (g) 2.0 (14 wt %) 1.5 (14 wt %) (wt % ofproteinaceous matter) Total isoleucine (g) 1.0 0.75 Total valine (g) 1.00.75 EAA (g) 7.0 (50 wt %) 5.3 (50 wt %) (wt % of proteinaceous matter)Fat (g) 2.0 1.5 Digestible carbohydrates (g) 6.4 4.8 Indigestiblecarbohydrates (g) 1.11 0.83 GOS (g) 0.83 0.63 FOS/inulin (g) 0.09 0.07Low-viscosity pectin (g) 0.19 0.14 Ca (mg) 332 250 Mg (mg) 25 19 Fe (mg)1.6 1.2 Zn (mg) 1.5 1.1 Se (μg) 10 7.5 Carotenoids (μg) 200 150 VitaminC (mg) 21.3 16.0 Vitamin E (mg-α-TE) 5.0 3.8 Vitamin D3 (μg) 13.3 10.0Vitamin B6 (μg) 500 375 Folic acid (μg) 133 100 Vitamin B12 (μg) 2.0 1.5pH 4

EXEMPLARY EMBODIMENTS OF THE PRESENT INVENTION Embodiment (A)

A sterilized liquid acid enteral nutritional composition comprising per100 ml of said composition 9 to 20 g of non-hydrolysed globular protein,fat, and at least 100 mg of divalent metal cations and having a pHranging between 3 and 5.

Embodiment (B)

Nutritional composition according to the preceding embodiment, whereinthe pH ranges between 3.7 and 4.3, preferably is equal to about 4.0.

Embodiment (C)

Nutritional composition according to any one of the precedingembodiments, wherein the amount of divalent metal cations ranges between100 and 600 mg per 100 ml.

Embodiment (D)

Nutritional composition according to any one of the precedingembodiments, wherein the amount of non-hydrolysed globular proteinranges between 4 and 16 g, preferably is equal to about 10 g per 100 mlof the composition.

Embodiment (E)

Nutritional composition according to any one of the precedingembodiments, wherein the divalent metal cation is selected from thegroup consisting of Ca, Mg and any mixture thereof, preferably is Ca.

Embodiment (F)

Nutritional composition according to any one of the precedingembodiments, wherein the globular protein is selected from the groupconsisting of whey protein, pea protein, soy protein, and any mixturethereof.

Embodiment (G)

Nutritional composition according to embodiment 6, wherein the source ofwhey protein is selected from the group consisting of whey proteinconcentrate (WPC), whey protein isolate (WPI), and any mixture thereof.

Embodiment (H)

Nutritional composition according to any one of the precedingembodiments, wherein the amount of non-hydrolysed globular protein is atleast 85 weight % of the total proteinaceous matter in the composition.

Embodiment (I)

Nutritional composition according to any one of the precedingembodiments, further comprising a non-globular protein, a hydrolysedprotein, an oligopeptide, a peptide or a free amino acid.

Embodiment (J)

Nutritional composition according to embodiment 9, wherein thenon-globular protein is selected from the group of casein, caseinate,micellar casein isolate, and any mixture thereof.

Embodiment (K)

Nutritional composition according to embodiment 9, wherein the freeamino acid is selected from the group of branched chain amino acids, inparticular is L-leucine.

Embodiment (L)

Nutritional composition according to any one of the precedingembodiments, said fat providing between 15 to 65% of the total energyamount of the composition.

Embodiment (M)

Nutritional composition according to any one of the precedingembodiments, further comprising carbohydrate, said carbohydrateproviding between 20 to 60% of the total energy amount of thecomposition.

Embodiment (N)

Nutritional composition according to any one of the precedingembodiments, wherein the viscosity of the composition is lower than 200mPa·s, preferably lower than 100 mPa·s, measured at 20° C. at a shearrate of 100 s⁻¹.

Embodiment (O)

Nutritional composition according to any one of the precedingembodiments, in a unit dosage container of about 200 ml.

Embodiment (P)

Nutritional composition according to any one of the precedingembodiments comprising:

-   a) about 10 g of non-hydrolysed whey per 100 ml of the composition,    said protein providing about 56% of the total energy amount of the    composition;-   b) fat providing about 18% of the total energy amount of the    composition;-   c) optionally carbohydrate providing about 23% of the total energy    amount of the composition,-   d) about 250 mg per 100 ml of Ca and about 19 mg of Mg per 100 ml;    and-   e) having a pH of about 4.

Embodiment (Q)

The use of a nutritional composition according to any one of thepreceding embodiments for the manufacture of a nutritionally completefood.

Embodiment (R)

Nutritional composition according to any one of the precedingembodiments, for use in nutritional management of a person in needthereof.

Embodiment (S)

Nutritional composition according to embodiment (R), wherein the personis an elderly person, a person that is in a disease state, a person thatis recovering from a disease state, a person that is malnourished, asportsman, or an active elderly.

Embodiment (T)

Nutritional composition according to embodiment (R) or (S) for theprevention or treatment of a disease or condition involving muscledecline in a mammal, in particular for treating sarcopenia.

Embodiment (U)

A method for the preparation of a sterilized liquid acid enteralcomposition comprising per 100 ml 9 to 20 g of non-hydrolysed globularproteins, fat and at least 100 mg of divalent metal cations and having apH ranging between 3 and 5 according to any one of the embodiments (A)to (P), comprising a step wherein at least the non-hydrolysed globularproteins are subjected to a homogenization step, followed by directsteam injection (DSI) at specific holding values, such as a holdingtemperature of 100 to 140° C. during a holding time of about 0.5 to 10seconds, followed by a sterilization step.

Embodiment (V)

Method according to embodiment (U), comprising the consecutive steps of:

-   a) preparing an aqueous solution comprising amounts of divalent    metal cations, in particular calcium and magnesium, non-hydrolysed    globular proteins and fat, such that said sterilized liquid acid    enteral composition comprises per 100 ml of said composition 9 to 20    g of non-hydrolysed globular proteins, fat, and at least 100 mg of    divalent metal cations, and having a pH ranging between 3 and 5;-   b) homogenizing the resulting solution essentially obtained by step    a);-   c) subjecting the resulting solution essentially obtained by step b)    to a direct steam injection process at a holding temperature of 100    to 140° C. during a holding time of about 0.5 to 10 seconds; and-   d) subjecting the resulting solution essentially obtained by step c)    to a sterilization treatment.

Embodiment (W)

Sterilized liquid acid enteral composition comprising per 100 ml 9 to 20g of non-hydrolysed globular proteins, fat and at least 100 mg ofdivalent metal cations and having a pH ranging between 3 and 5,obtainable by a method according to embodiment (U) or (V).

1. A sterilized liquid or semi-solid acid enteral nutritionalcomposition comprising per 100 ml of said composition 9 to 20 g ofnon-hydrolysed globular protein, fat, and at least 100 mg of divalentmetal cations and having a pH ranging between 3 and
 5. 2. Thenutritional composition according to claim 1, wherein the pH rangesbetween 3.7 and 4.3.
 3. The nutritional composition according to claim1, further comprising a stabilizing polysaccharide selected from thegroup consisting of (high) methoxy pectin and carboxymethyl celluloseand combinations thereof.
 4. The nutritional composition according toclaim 1, wherein the amount of divalent metal cations ranges between 100and 600 mg per 100 ml.
 5. The nutritional composition according to claim1, wherein the amount of non-hydrolysed globular protein ranges between9 and 16 g.
 6. The nutritional composition according to claim 1, whereinthe divalent metal cation is selected from the group consisting of Caand Mg and any mixture thereof.
 7. The nutritional composition accordingto claim 6, wherein the globular protein is selected from the groupconsisting of whey protein, pea protein, soy protein, and any mixturethereof.
 8. The nutritional composition according to claim 1, whereinthe globular protein comprises whey protein and the source of wheyprotein is selected from the group consisting of whey proteinconcentrate (WPC), whey protein isolate (WPI), and any mixture thereof.9. The nutritional composition according to claim 1, wherein the amountof non-hydrolysed globular protein is at least 85 weight % of the totalproteinaceous matter in the composition.
 10. The nutritional compositionaccording to claim 1, further comprising free amino acid.
 11. Thenutritional composition according to claim 1, said fat providing between15 to 65% of the total energy amount of the composition.
 12. Thenutritional composition according to claim 1, further comprisingcarbohydrate, said carbohydrate providing between 20 to 60% of the totalenergy amount of the composition.
 13. The nutritional compositionaccording to claim 1, wherein the composition is a liquid and theviscosity of the composition is lower than 100 mPa·s, measured at 20° C.at a shear rate of 100 s⁻¹. 14-27. (canceled)
 28. The nutritionalcomposition according to claim 1, wherein the pH is equal to about 4.0.29. The nutritional composition according to claim 1, wherein the amountof non-hydrolysed globular protein is equal to about 10 g per 100 ml ofthe composition.
 30. The nutritional composition according to claim 1further comprising free branched chain amino acid.